bioscience price list 2014-15 - volume i

8/16/2019 Bioscience Price List 2014-15 - Volume I

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9BioscienceAvailability of the products that of animal origin is subject to government regulations.Please check with your local MM representative before ordering. www.merckmillipore.com

Genomic Products

Proteomic Products

Custom Research Services & Products

Educational Products

Techware Equipments

New Products

GENEI


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Part 1

Genei

10 Bioscience Availability of the products that of animal origin is subject to government regulations.Please check with your local MM representative before ordering.www.merckmillipore.com

Genei restriction enzymes are supplied in convenient pack sizesand suitable concentrations. We are supplying commonly usedenzymes in high concentration to aid genomic DNA digestions.Each lot of enzyme is rigorously checked for its integrity andfunctional purity during and after the process. All batches ofrestriction enzymes are assayed for their activity once everytwo months. Stability checks have confirmed that most ofthe enzymes are stable at -20° C for more than one year.Concentrated enzymes stay active longer than dilute enzymes.

We have divided the restriction enzymes based on the assayconditions into five groups for the convenience of the user.The enzymes and the buffer vials are colour coded. 10X assay

buffer and 100X nuclease free BSA (wherever needed) is suppliedfree of cost with the enzyme. Genei also caters to your additionalrequirement of reaction buffers. For reaction buffer set referindex. Unique buffer is supplied for the enzyme which performssub-optimally with the existing five assay buffers. The detailsof incubation buffer system is given in the catalogue.

Quality Checks:

Unit Definition:

The catalytic activity of the restriction enzymes available fromus is based on the determination of the minimum amount ofenzyme required for the generation of the enzyme-specific final

fragment pattern of lambda DNA in most cases.One unit is defined as the amount of enzyme required to producea complete digest of 1 µg of lambda DNA in a reaction volumeof 50 µl in 60 minutes under optimal conditions of salt, pHand temperature.

In some cases the determination of the unit is based on digestionof λ dam- DNA, pBR 322 DNA, Ad2 DNA,λ / EcoR I digest orλ / Hind III digest.

Overnight Non-specific Nuclease Assay:

Every preparation of restriction enzyme is tested for non-specific nucleases. Varying units of enzyme is incubated with

1 µg of substrate DNA at the recommended assay conditionsin 50 µl reaction volume for 16 - 20 hours. A sharp unalteredbanding pattern is an indication of the purity of enzyme freefrom detectable non-specific nucleases. The highest numberof units producing sharp unaltered pattern is reported on thecertificate of analysis supplied with each enzyme.

Non-Specific Endonuclease assay:

The test is done for enzymes that do not have any site insupercoiled plasmid substrate. These enzymes are incubatedwith 1 µg of supercoiled (RFI form) DNA in 0.05 ml reaction

Restriction Enzymes

volume for 4 hours at the recommended assay conditions. Asingle non-specific nick in the supercoiled form produces nicked(RF-II) form. The two forms can be distinguished on an agarosegel and the percentage of conversion estimated.

Ligation/Recut Assay:

The DNA fragments produced by a 3 to 10 fold excess ofrestriction enzyme digestion are ligated. Following the ligation,the DNA fragments are recut with the same restriction enzyme.An unaltered banding pattern after cleavage indicates intact5’ and 3’ termini as well as the absence of contaminatingnucleases. The estimated level of ligation and re-cleavage mustexceed 70-90%.

Blue/White Standard Assay:

This is a very sensitive assay for the restriction enzymes used incloning applications. This assay allows the detection of very lowlevels of nuclease contamination present in restriction enzymes.This assay is performed for those enzymes that have site presentin the multiple cloning region that is in turn present within LacZα gene of the plasmid.

The assay is performed by cleaving the plasmid with 3-10fold excess of enzyme. The cleaved DNA is ligated and usedto transform competent DH5α E.coli cells. The transformedcells are grown on a selective medium to ensure that all the

colonies that grow arise from an E.coli transformed with theligated vector. These colonies are tested for the integrity of thepolylinker site by checking for the presence of the functionalß-galactosidase locus. Only those colonies that turn blue onaddition of X-gal and IPTG have ß-galactosidase locus intactthat in turn indicates that the polylinker site (that lies withinthis structural gene) was unchanged by cut and religation. Thepermitted percentage of white colonies is upto 2% for stickyend generating enzymes and upto 10% for blunt end generatingenzymes.

Star activity: Genei recommends very careful use of a fewenzymes which are very sensitive to slightly altered assayconditions. The enzymes which exhibit star activity when highunits/µg of DNA and/or incubated for long hours are BamH I,EcoR I, Kpn I, Nco I, Hinf I, PvuII, Sau3A I, Ssp I, Sal I, Nhe Iand Kpn I.

Genomic


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Tips to use Restriction Enzymes

• All restriction enzymes and 10X assay buffers should be storedat -20°C. Thaw the assay buffers completely before use.

• Restriction enzymes should be kept on ice when they are notin the freezer.

• The enzyme should always be the last component added tothe reaction mixture.

• The substrate DNA should be free of contaminations such asphenol, chloroform, alcohol, EDTA, detergents or excessivesalts all of which can interfere with restriction enzymeactivity.

• Nature of DNA and DNA Methylation is also an importantelement of a restriction digestion so it is advisable to checkthe amount of enzyme needed to cleave the substrate priorto the actual experiment.

• The restriction enzyme: DNA: reaction volume ratio according

to the unit definition is 1 U: 1 µg: 50 µl and can be used asa guide when designing a reaction mixture.

• Smaller volumes are more susceptible to pipetting errors.• An important point to keep in mind for a successful digestion

is mixing. Ensure thorough mixing for complete digestion. Werecommend gentle up and down pipetting of reaction mixturefollowed by a short spin in microcentrifuge.

• Incubation time may be shortened if an excess of restrictionenzyme is added to the reaction mixture and vice versa.

• Enzymes should be diluted (if required) using respectivedilution buffer. The diluted enzymes should be used thesame day.

•

The termination of the reaction may be done either by addingstop solution or by heat inactivation at 65°C for 20 minutes.Phenol-chloroform extraction may be followed as analternative method for the restriction enzymes that cannotbe heat inactivated.

Factors Influencing RestrictionEnzyme Activity Nature of DNA :The nature of the substrate strongly influences the activity ofrestriction enzymes. The most important parameters are :

• base distribution in natural DNA•

tertiary structure of DNA• base composition of the flanking sequence• the position of the cleavage site with respect to each other.

If the DNA has contaminants like phenol, chloroform, alcohol,detergents, EDTA the restriction enzyme activity will beinhibited.

Temperature :

Optimal digestion of DNA varies over a wide temperaturerange for different restriction enzymes. The restrictionenzymes supplied by Genei include enzymes for which

the optimum incubation temperature is d ifferentfrom the standard incubation temperature of 37°C.These enzymes are listed below.

Enzymes Assay Temperature

Apa I 25°C

Bst E II 60°C

Sma I 25°C

Taq I 65°C

Tris-HCl is the most commonly used buffering agent in incubationmixtures. This buffer system is markedly temperature dependent.The change in pH per 10°C amounts to approx 0.3.

Another important factor affecting the optimum activity isthe appropriate ionic environment. Mg2+ ions are an absoluterequirement for all restriction enzymes, whereas the addition

of other salt components depends on the different nucleases.Sometimes the presence of BSA in the reaction mix has thecrucial influence on the activity of enzymes, because it stabilizesthe enzyme, binds some impurities, prevents the enzymeadsorption to the test tube surface.

Factors Influencing RestrictionEnzyme Activity contd...Methylation of DNA :

Restriction endonucleases are part of prokaryotic restriction/modification systems. The digestion of DNA isolated duringcloning steps in bacterial cells can be strongly affected by themethylation of specific adenosine or cytosine residues in therecognition sequence of the restriction enzyme of interest.

Many E.coli host strains possess two nucleotide sequence specificmethylases; thedam methylase which modifies adenine residues toN⁶-methyladenine in the sequence GATC and the dcmmethylase which modifies the internal cytosine residues to5-methylcytosine in CCAGG or CCTGG sequences. Digestion ofDNA may be inhibited by dam ordcm methylation sequences.

Restriction enzymes affected by dam anddcm methylation arelisted below:

Enzymes inhibited

by dam methylation

Enzymes inhibited

by dcm methylation

Cla I

+

AT/CGAT

+

Stu I AGG/CCT

Mbo II

+

GAAGA(N)8/7

Mbo I

+

/GATC

Nru I

+

TCG/CGA

Taq I

+

T/CGA

Xba I

+

T/CTAGA


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Part 1

Genei


Reaction Buffer for Restriction EnzymesGenei provides colour coded10X assay buffer with each restriction enzyme to ensure optimal activity. Some restriction enzymesrequire BSA at a final concentration of 100 µg/ml for optimal activity. BSA is supplied as 10 mg/ml (100 X) stock when requiredand should be added to the reaction mixture.

Final Concentration in mM (1X Recipe)

Buffer Tris HCl Tris -

Acetate

Sodium

Chloride

Magnesium

Chloride

Magnesium

Acetate

Potassium

Acetate

Potassium

Chloride

DTT pH Enzymes

A 10 — 150 7 — — — 1 7.9 EcoR V, Not I*, Sal I

B 10 — 100 10 — — — 1 8.0 Bgl I, Bgl II, Hinc II,

Mlu I, Ssp I.

C 10 — 50 10 — — — 1 7.8 Alu I, Hae III, Hinf I, Msp I,

Nhe I, Pvu II,StuI, Xba I.

D Optimised

Conc.

— — 10 — — Optimised

Conc.

— Optimised

pH

BamH I, BstE II, EcoR I,

Hind III, Mbo I, Mlu I, Nsi I,

Taq I, Pst I, Pvu I, Spe I, Nco

I, Nde I, Nru I

L 10 — — 10 — — — 1 7.4 Hpa II, Sac I, Xma I

E — 33 — — 10 66 — 0.5 7.9 Apa I, Ava I, Cla I, DraI, Hpa

I, Nae I, Sau3A I, SnaB I, Sfi

I, Sma I, Xho I, Xmn I.

Note :

Not I* Buffer A with 0.01% Triton X 100. The enzymes printed bold need BSA for optimum activity.

CpG Methylation is found in higher eukaryotes. These enzymes(e.g. Dnmt1) transfer a methyl group to the C5 position ofcytosine residues. Patterns of CpG methylation are heritable,tissue specific, and correlate with gene expression. ConsequentlyCpG methylation has been postulated to play a role indifferentiation and gene expression.

This CpG methylation patterns will not be retained once theDNA is cloned into a bacterial host.

Genei Restriction Enzymes inhibited by CpG methyltion arelisted below:

Enzymes inhibited by CpG methylation

Ava I C/PyCGPuG

Cla I AT/CGAT

Hha I GCG/C

Hpa II C/CGG

Mlu I A/CGCGT

Nae I GCC/GGC

Nru I TCG/CGA

Pvu I CGAT/CG

Sal I G/TCGAC

Sfi I GGCCNNN/NGGCC

Sau3AI /GATC

Sma I CCC/GGG

Xma I GAANN/NNTTC


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List of IsozymesSl. No. Restriction Enzymes Sequence Isozymes

1. Apa I GGG CC↓C Bsp120 I, PspOM I

2. Ava I C↓PyCGPuG Eco88 I

3. BstE II G↓GTNACC BstP I, Eco91 I, Eco065 I, PspE I

4. Cla I AT↓CGAT Ban III, Bsa29 I, Bsc I, BseC I, BsiX I, Bsp106 I, BspD I, BspX I, Bsu15 I

5. EcoR V GAT↓ATC Eco32 I

6. Hae III GG↓CC Bsh I, BsuR I, Pal I

7. Hinc II GT(T/C)↓(A/G)AC Hind II

8. Hpa I GTT↓AAC BsHP I, KspA I

9. Hpa II C↓CGG Bsi S I, Hap II, Msp I

10. Kpn I GGTAC↓C Acc65 I, Asp718

11. Mbo I ↓GATC BscF I, Bsp143 I, Dpn II, Kz09 I, Nde II, Sau3A I

12. Msp I C↓CGG Bsi S I, Hap II, Hpa II

13. Nae I GCC↓GGC MroN I, NgoA IV, NgoM IV

14. Nco I C↓CATGG Bsp19 I

15. Nde I CA↓TATG FauND I

16. Not I GC↓GGCCGC CciN I

17. Nhe I G↓CTAGC AsuNH I

18. Nsi I ATGCA↓T EcoT22 I, Mph1103 I, Ppu10 I, Zsp2 I

19. Nru I TCG↓CGA Bsp68 I

20. Pvu I CGAT↓CG BspC I, Ple19 I

21. Sac I GAGCT↓C Ecl136 II, Eco ICR I, Psp124B I, Sst I

22. Sau3A I ↓GATC BscF I, Bsp143 I, Dpn II, Kzo9 I, Mbo I, Nde II

23. Sma I CCC↓GGG Cfr9 I, PspA I, Xma I, XmaC I

24. SnaB I TAC↓GTA BstSN I, Eco105 I25. Spe I A↓CTAGT AclN I, Bcu I

26. Stu I AGG↓CCT Aat I, Eco147 I, Pme55 I, SseB I

27. Taq I T↓CGA TthHB8 I

28. Xho I C↓TCGAG PaeR7 I, Sfr274 I, Tli I

29. Xma I C↓CCGGG Cfr9 I, PspA I, Sma I, XmaC I

30. Xmn I GAANN↓NNTTC Asp700 I, MroX I


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Relative Activity of Restriction Enzymes in Genei Assay Buffer SystemSl. No. Restriction

Enzymes

10XAssay Buffers

A B C D E L

1. Alu I 50 75 100 100 100 1002. BamH I 25 50 50 100 50 50

3. Bgl I 100 100 75 100 25 0

4. Bst E II 0 50 25 100 50 0

5. Eco R I 25 50 50 100 50 0

6. Eco R V 100 75 50 100 50 0

7. Hae III 75 100 100 100 100 25

8. Hpa I 0 0 0 25 100 25

9. Kpn I 0 0 75 25 50 100

10. Mbo I 50 75 50 100 50 0

11. Msp I 25 75 100 25 100 100

12. Nco I 0 50 50 100 50 50

13. Not I 100 100 75 100 0 0

14. Pst I 50 50 50 100 25 5015. Sac I 25 25 50 50 100 100

16. Sal I 100 50 0 25 0 0

17. Sau 3A I 25 25 25 100 100 75

18. Spe I 25 25 50 100 50 50

NOTE:

• supplies the buffers that are typed in italics and bold with the respective enzymes. • Chart serves as a guide to choose the compatible buffer for double digestion. • In case of Taq I, the percentage activity is reported with respect to the unique buffer supplied.

Cat No. PI. No. Product Sequence Assay Condition Volume

Activity

(Units/µl).

Qty Unit Price

` Buffer Temp

610101700041730 MBE17L Alu I AG↓CT C 37°C 5-10 500 U 6000

610100100021730 MBE1S BamH I G↓GATCC D+ BSA 37°C 10 2000 U 1250

610100300021730

610100300041730

MBE3S

MBE3L

EcoR I

High Conc.

G↓AATTC D+ BSA 37°C 20

20

4000 U

20000 U

1040

3600

610100900021730 MBE9S EcoR V GAT↓ATC A+ BSA 37°C 10 1200 U 1800

610101000041730 MBE10L Hae III GG↓CC C 37°C 10 4000 U 5060

610100600041730 MBE6L Hind III A↓AGCTT D+ BSA 37°C 20 4000 U 1710

610102200041730 MBE22L Kpn I GGTAC↓C L+BSA 37°C 10 4000 U 4200

610102700041730 MBE27L Mbo I ↓GATC D+BSA 37°C 5-10 400 U 5780

610103100021730 MBE31S Msp I C↓CGG C 37°C 10 400 U 1790

610101500041730 MBE15L Nco I C↓CATGG D+ BSA 37°C 5-10 400 U 5750

610106200021730 MBE62S Nde I CA↓TATG D+ BSA 37°C 20 4000 U 1530

610101600041730 MBE16L Not I GC↓GGCCGC A+ BSA 37°C 5-10 400 U 5560

610101200021730 MBE12S Pst I CTGCA↓G D + BSA 37°C 10 1200 U 1960

610101800041730 MBE18L Sac I GAGCT↓C L+BSA 37°C 10 2000 U 6090

610100400041730 MBE4L Sal I G↓TCGAC A+BSA 37°C 10 2000 U 4030

610100500041730 MBE5L Sau3A I ↓GATC E+BSA 37°C 5-10 400 U 6000

610103800041730 MBE38L Spe I A↓CTAGT D+BSA 37°C 1-5 400 U 5340

610100700021730 MBE7S Taq I T↓CGA Unique 65°C 10 800 U 2320

All restriction enzymes and 10X assay buffers should be stored at -20°C


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Modifying Enzymes

Quality Control for Modifying Enzymes

Unit definitions

Unit definition varies from enzyme to enzyme and is given inthe product description of this catalogue.

Performance Test

Each enzyme is tested for its performance using tests that arementioned in product description.

Assay for non-specific endonuclease

All modifying enzymes are checked for the absence ofnon-specific endonuclease activity by incubating excess of theenzyme with supercoiled plasmid DNA for several hours anddetermining the level of nicking in supercoiled form. The highestnumber of units producing unaltered pattern is reported on the

data sheet supplied with the enzyme.

Assay for exonuclease activity

The absence of exonuclease activity is checked for all theenzymes by incubating the enzyme with lambda substrate for

several hours and running on an agarose gel. The sharp patternindicates the absence of exonuclease. The highest number ofunits producing unaltered pattern is reported on the data sheetsupplied with the enzyme.

Assay for RNase

The absence of contaminating RNases is tested by incubatingtotal RNA from HeLa cells with excess of enzyme for 4 hoursat 37°C and checking for any degradation on gel.

Purification

All modifying enzymes are purified to near homogenity usingthe procedures developed at Genei.

Stability

All batches of modifying enzymes are routinely tested for

stability by checking unit activity. Most of the enzymes arestable for more than 12 months when stored at -20°C. Exposureto temperatures greater than -20°C should be minimisedwhenever possible.

T4 DNA Ligase

T4 DNA ligase catalyses the linkage of adjacent 5'-phosphate

and 3'-hydroxy ends of double stranded DNA by the formationof phosphodiester bond.

Unit Definition : One cohesive end ligase unit is defined as theamount of enzyme required to give 50% ligation of Lambda/Hind III digest in 30 minutes at 16°C in 20 µl of the reactionmixture and a 5' DNA termini concentration of 0.12 µM.

Relation to other Ligase Units : One cohesive end Ligase unitapproximately equals 0.015 ATP-PP exchange unit (Weiss Unit).

Storage Buffer : 10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 50 mMKCl, 1 mM DTT, 50% Glycerol and 200 µg/ml Nuclease free BSA.

Storage : -20°C

Note :

T4 DNA Ligase is supplied with 1 vial each of Cohesive & Bluntend Buffer.

For Buffer Composition refer Buffer Set for Ligation given onpage no C14.

Application : This enzyme seals nicks in double stranded DNAand covalently joins DNA fragments with cohesive ends or bluntends. This enzyme does not act on single stranded DNA.

Performance Test :

1. T4 DNA Ligase is routinely used in-house for qualitycontrol checks of all our cohesive and blunt end generatingrestriction enzymes. It is also used in the Blue-Whitestandard assay.

2. Enzyme is tested for its performance in cloning experiments.

Blunt End Ligation

Ligation of λHae III blunt ended fragments using 1 µl of T4DNA ligase at 22°C at different time points from 0-60 minutes.

Ordering Information :

Cat # PI. # Product Unit Price

`

611100100021730 MME1S T4 DNA Ligase

(8000 U)

2200

611100100041730 MME1L T4 DNA Ligase

(40000 U)

5700

611160100021730 MME1HC T4 DNA Ligase (High

Conc.) (100000 U)

12080


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Part 1

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INSTANT Ligation KitThe INSTANT Ligation Kit is specially designed for efficientligation of both cohesive and blunt ended DNA fragments in

just 5 minutes at room temperature (25°C). The kit consists of

INSTANT T4 DNA ligase and 2X INSTANT ligation buffer thatsignificantly speeds up the process of ligation compared toovernight incubation. The ligated products can be directly usedfor transformation without any further purification steps.

This kit is supplied with reagents sufficient to perform 20reactions.

Application :

• Circularization of linear DNA• Cloning DNA fragments into vector• Construction of genomic library• TA Cloning

Highlights :

• Ligation of both blunt and cohesive ends using same buffer.• Incubation for just 5 minutes at room temperature (25°C).• No additional purification steps required prior to

transformation.

Storage : -20°C



`

611660300011730 KT91 Instant Ligation Kit (20

Reactions)

3810

Buffer Set for LigationFollowing Buffers can be ordered from us.

Assay buffers :

Cohesive End Ligation Buffer (1X) : 50 mM Tris-HCl (pH 7.4),10 mM MgCl₂, 20 mM DTT, 50 µg/ml Nuclease free BSA and1 mM ATP.

Buffer supplied at 10X concentration.

Blunt End Ligation Buffer (1X) : 50 mM Tris - HCl (pH 7.4),10 mM MgCl₂, 1 mM DTT, and 1mM ATP, 5% w/u PEG.

Buffer supplied in 5X concentration.

Storage : -20°C



`

611150300011730 MELB3 Cohesive & Blunt End Buf-

fer (0.5 ml ea.)

1860

611150400011730 MELB4 Dilution Buffer for T4 DNA

Ligase (0.5 ml x 2)

1760

T4 Polynucleotide KinaseT4 Polynucleotide Kinase catalyses the transfer of gammaphosphate group from ATP to the 5'-hydroxy group ofpolynucleotides.

Unit Definition : One unit is defined as the amount of enzymecatalysing the production of one nmole of acid insoluble 32Pin 30 minutes at 37°C.

Assay Buffer (1X) : 70 mM Tris-HCl (pH 7.6), 10 mM MgCl₂and 5 mM DTT.


Storage Buffer : 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mMDTT, 50 mM KCl, 50% Glycerol and 200 µg/ml Nuclease free BSA.

Application : T4 Polynucleotide Kinase is very useful in the 5'end labelling of nucleic acids.

Performance Test : Labelling and Kination efficiencies areevaluated.

Storage : -20°C



`

611100400051730 MME4L T4 Polynucleotide

Kinase 1000 U

5870

Alkaline Phosphatase (Calf IntestinalPhosphatase) (Molecular Biology Grade)

Alkaline phosphatase molecular biology grade catalyzes the

removal of 5'-phosphate groups from DNA, RNA and ribo - anddeoxyribonucleoside triphosphates.

Unit Definition : One unit is defined as the amount of enzymethat hydrolyses 1 µ mole of p-nitrophenyl phosphate top-nitrophenol in 1 minute at 37°C in a volume of 1 ml.

Assay buffer (1X) : 50 mM NaCl, 10 mM Tris-HCl (pH 7.9),10 mM MgCl₂ and 1 mM DTT.

Buffer Supplied at 10X concentration.

Storage buffer : 50 mM KCl, 10 mM Tris-HCl (pH 8.2), 1 mMMgCl₂, 0.1 mM ZnCl₂ and 50% glycerol.

Application : CIP is used to dephosphorylate vectors in cloning

experiments to prevent vector self ligation and to preparetemplates for 5' end labelling.

Performance Test : Dephosphorylation efficiency is evaluated.

Storage : 4°C



`

611100800031730 MME8S Alkaline Phosphatase

(CIP) (Molecular Biology

Grade), 200 Units

2450


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DNase I (RNase Free)DNase I (bovine pancreas) is a glycoprotein and double strandspecific endonuclease which is made free of RNase activity. Itis best suited for specific applications, where maintenance of

RNA integrity is critical. The protein requires divalent cationsfor maximal activity. Supplied in 20mM Tris-HCl (pH 7.6), 1mMDTT, 0.1mg/ml Nuclease Free BSA and 50% (v/v) glycerol.

Unit Definition : One unit is that amount of enzyme causing anincrease in absorbance at 260 nm by 0.001 per minute at 25°Cand pH 5.0.

Application :

DNase I (RNase free) is used in purification of DNA - free RNAmade by in-vitro synthesis using SP6 or T7 RNA Polymerasesystem.

Used for radioactive labelling by nick translation.Storage : -20°C.



`

610655600011730 FC56 DNase I (RNAse Free)

1000 U

8550

Deoxyribonuclease ISupplied as white lyophilised powder which dissolves readily at5 mg/ml in 0.15 M sodium chloride/glass distilled water to

give a clear colourless solution. The enzyme loses its activityirreversibly by heat treatment at 80°C for 10 min.

Activity : In the range of 1000 U/mg material (Kunitz).

Unit definition : One unit is that amount of enzyme causingan increase in absorbance at 260 nm by 0.001 per min. at 25°Cand pH 5.0.

Storage : -20°C

Application : Deoxyribonuclease I (bovine pancreas) is usedto catalyze random degradation of both single and double-stranded DNA producing 5'-P terminal oligonucleotides.



`

612150280501730 FC28L Deoxyribo-nuclease

I 50 mg

5780

RNase A (Ribonuclease A)Ribonuclease A (bovine pancreas) supplied as white lyophilizedpowder which dissolves readily at concentration of 2 mg/ml inanalytical grade water to give a clear colourless solution.

Purity : > 70% pure as checked by ion exchange chromatography.

Activity : In the range of 70 - 80 U/mg protein.

Enzyme is not tested for DNase activity.

Unit definition:One unit is that amount of enzyme causing hydrolysisof RNA at a rate such that k (velocity constant) equals unity at25°C and pH 5.0.

Storage : -20°C

Application :

Used in phage extraction



`

612150380501730 FC34S RNase A 50 mg 2380

612150382501730 FC34L RNase A 250 mg 8360

RNase A (Ribonuclease A) (DNase Free)Ribonuclease A (bovine pancreas) is used to hydrolyse RNAduring isolation of DNA. Supplied as lyophilized powder whichdissolves readily in analytical grade water.

Essentially DNase free and > 95% pure as checked by ionexchange chromatography.

Activity : In the range of 80 - 90 u/mg protein.

Unit definition: One unit is that amount of enzyme causing hydrolysisof RNA at a rate such that k (velocity constant) equals unity at

25°C and pH 5.0.

Storage : -20°C

Application :

• In the isolation of DNA for RNA-free DNA.• As a molecular weight marker.

Performance Test : Efficiency tested in plasmid preparationby alkali lysis.

DNase is not detected.

Absence of DNase contamination : Varying units of RNase A(DNase Free) is incubated with 1 µg of substrate DNA at the

recommended assay conditions in 50 µl reaction volume for4 hours. A sharp unaltered banding pattern is an indicationof the purity of RNase A (DNase Free) free from detectablenon-specific nucleases.



`

612150480101730 FC25S RNase A (DNase Free) 10 mg 1510

612150480501730 FC25L RNase A (DNase Free) 50 mg 4800

612150481001730 FC25J RNase A (DNase Free) 100 mg 8020


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Part 1

Genei


Human Placental RNase InhibitorHuman Placental RNase Inhibitor is an acidic protein ofmolecular weight near 50 Kda. It forms 1:1 complex withRNase A and is a noncompetitive inhibitor of this enzyme.

RNase Inhibitor is active over a broad pH range and requiresDTT for its activity.

Unit definition : One unit is defined as the amount ofRNase Inhibitor required to inhibit the activity of 5 ng of RNaseA by 50%.

Storage buffer : 20 mM Hepes-KOH (pH 7.6), 50 mM KCl, 5 mMDTT and 50% glycerol.

Application : RNase Inhibitor is used to protect the mRNA incDNA synthesis, in in vitro transcription /translation system andin vitro RNA synthesis.

Note : 5 mM DTT concentration is critical for RNase Inhibitorand so has to be maintained during long and repeated uses.

Storage : -20°C

Performance Test : Used extensively in our lab for in vitrotranscription assays and RT-PCRs.



`

611101100011730 FC11 Human Placental RNase

inhibitor 1000 U

5800

Recombinant RNase InhibitorRecombinant RNase Inhibitor is the recombinant form ofHuman Placental RNase Inhibitor. The complete ORF of thegene is cloned and expressed in E coli , hence retaining identicalproperties and performance of its native form. This Productcan be used for the same applications as those from the nativeform of the protein.

Unit definition : One unit is defined as the amount of RNaseInhibitor required to inhibit the activity of 5 ng of RNase A by50%.

Storage buffer : 20 mM HEPES-KOH (pH 7.6), 50 mM KCl,

5 mM DTT and 50% glycerol.

Application : RNase Inhibitor is used to protect the mRNA incDNA synthesis, in in vitro transcription /translation system andin vitro RNA synthesis.

Note : 5 mM DTT concentration is critical for RNase Inhibitorand so has to be maintained during long and repeated uses.

Storage : -20°C

Performance Test : Used extensively in our lab for in vitrotranscription assays and RT-PCRs.



`

611101200011730 FC73S Recombinant RNase

inhibitor,1000 Units

3940

Products for PCRPolymerase Chain Reaction (PCR) is an effective tool tosynthesize defined sequences of DNA using thermostablepolymerases. PCR exploits the inherent stability of theseenzymes at high temperatures to amplify about a million copiesof desired DNA fragment using two specific oligonucleotideprimers that anneal to opposite strands flanking the target DNAsequence to be amplified.

PCR is now widely used in cloning and sequencing DNAfragments, as a diagnostic tool and to generate labelled probes.To cater to these varied applications, Genei has a range of PCRenzymes, buffers, kits and optimized protocols.

Taq DNA Polymerase : Taq DNA Polymerase, a thermostableenzyme used in most routine PCR amplifications for sizesranging from 100 bp to 3 Kb, is a highly pure enzyme that lacks3'-5' exonuclease (proof reading) activity. The enzyme can alsobe used in applications like RAPD, RFLP and AFLP studies todetect variations within the genomes of species.

HotStart Taq DNA Polymerase : HotStart Taq DNA Polymerase,used for improved PCR specificity, is an optimized mixture of

highly purified Taq DNA Polymerase with high affinity anti-Taqmonoclonal antibody that inhibits polymerase activity at suboptimal temperatures by binding to Taq DNA Polymerase.

Effi -Taq™ DNA Polymerase : Effi -Taq™ DNA Polymerase amodified form of Taq DNA Polymerase is supplied in an inactivestate that has no polymerase activity at ambient temperature.It is suitable for PCR systems with complex genomic templates,complex cDNA templates (RT-PCR), very low copy targets,multiple primers reaction (Multiplex PCR), high throughput PCRprocedures and systems that are prone to form Primer Dimer.

PR Polymerase : PR Polymerase, used in PCR reactions that

require high fidelity synthesis, is a thermostable enzyme thatexhibits strong ProofReading activity. The 3'-5' exonucleasemediated proofreading activity enables the enzyme torepair mis-incorporated nucleotides and reduce errorsduring polymerisation. This enzyme is ideal for high fidelityamplifications of short stretches of target DNA (< 1.5 kb).

An improved and more defined range of long PCR enzymeswith higher fidelity is now made available to the scientists. Forhigher sensitivity and better yield, we have two sets of longPCR enzyme systems.


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XT-5 PCR System : XT-5 PCR System, a unique enzyme blendthat utilizes the powerful processivity of Taq DNA polymeraseand the ProofReading activity of PR polymerase. This system isparticularly useful for amplifications of long target DNA fromgenomic DNA templates (guaranteed upto 5kb).

XT-20 PCR System : XT-20 PCR System, a unique enzyme mixoptimised for amplification of longer target DNA (guaranteedupto 20 kb) from genomic DNA templates.

Quality Control tests for Thermostable Polymerases.

Unit Definition :

One unit is defined as the amount of enzyme which incorporates10 nanomoles of total deoxyribo-nucleoside triphosphates intoacid precipitable DNA in 30 minutes at 72°C under optimalassay conditions.

Assay for non-specific endonuclease :

1 µg of supercoiled DNA incubated with excess enzyme at 37°Cand 72°C for 16 hours showed no relaxation of supercoiled DNAwhen visualised on a 1% agarose gel.

Assay for exonuclease activity :

1 µg of lambda DNA digest incubated with excess enzyme at37°C and 72°C for 16 hours showed no alteration in bandingpattern when visualised on a 1% agarose gel.

Performance test :

Performance tested as mentioned in product description ofeach enzyme.

Purification :

All thermostable polymerases are purified to near homogeneityusing procedures developed in-house.

Thermostability :

Taq DNA polymerase and PR polymerase are highly thermostableenzymes. They retain >90% activity even after prolongedincubation (1 hour) at 94°C.

Stability :

All batches of thermostable polymerases are periodically testedfor activity and are found to be stable for more than 12 monthswhen stored at -20°C. Exposure to temperature greater than

-20°C should be minimised.

Patents/Disclaimer :

Some products/processes mentioned in this catalogue may becovered by patents issued and applicable in certain countriesfor certain periods. Because purchase of our products does notinclude a licence to perform patented applications (in thesecountries) users have to ensure that they have the necessarylicence for use.

Products for PCR & RT-PCR

Enzymes Reagents Kits Master Mix

Taq DNA Polymerase

Red Taq DNA Polymerase

HotStart Taq DNA Polymerase

Effi-Taq DNA Polymerase

PR Polymerase

XT-5 PCR System

XT-20 PCR System

AMV Reverase Transcriptase

M-MuLV Reverse Transcriptase

dNTPS

10 mM Mix

100 mM Mix

10 mM Set

100 mM Set

Buffers

Taq Buffer A

Taq Buffer B

Taq Buffer E

Taq Buffer F

Taq Dilution Buffer

MgCl₂

MAGIC AmplificationSolution

DNA Amplification ReagentKit

PCR Master Mix Kit

Red Dye PCR Master Mix Kit

PCR Optimization Kit

Long PCR 5 kb Ampli Kit

Long PCR 20 kb Ampli Kit

Two Step AMV RT-PCR Kit

One Step AMV RT-PCR Kit

One Step M-MuLV RT-PCRKit

PCR Master Mix

Red Dye PCR Master Mix

HotStart PCR Master Mit


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T h e r m o p h i l i c P o

l y m e r a s e s S e l e c t i o n G u

i d e :

P r o d u c t

P r o d u c t s i z e

Y i e l d

S p e c i fi c i t y

F i d e l i t y

3 ’ - A

o v e r h a n g

E

a s e

G C r i c h

t e m p l a t e s

A p p l i c a t i o n

T a q D N A P o l y m e r a s e

U p t o 3 k b

●

●

●

● ●

●

●

R o u t i n e P C R

R A P D

R T - P C R

C o l o n y P C R

H i g h t h r o u g h

P u t P C R

P C R b a s e d D i a g n o s i s

M u l t i p l e x P C R

N o t e : T a q p o l a n d i t s v a r i a n t

c a n b e u s e d i n a l l t h e a b o v e

m e n t i o n e d a p

p l i c a t i o n s ,

c h o i c e i s b a s e

d o n t h e e n d

u s e r r e q u i r e m

e n t s f o r y i e l d ,

s p e c i fi c i t y e t c

. .

H o t S t a r t E n z y m e s a r e

p r e f e r r e d f o r S e n s i t i v e

a p p l i c a t i o n s

G e n e C l o n i n g

L o n g P C R L o n

g

R T - P C R

I n v i t r o M u t a g

e n e s i s

X T - P C R s y s t e m

s a r e

p r e f e r r e d f o r L o n g P C R

P C R M a s t e r M i x

U p t o 3 k b

●

●

●

● ●

● ●

●

R e d T a q D N A P o l y m e r a s e

U p t o 3 k b

●

●

●

● ●

● ●

●

R e d D y e P C R M a s t e r M i x

U p t o 3 k b

●

●

●

● ●

● ● ●

●

H o t S t a r t T a q D N A P o l y m e r a s e

U p t o 3 k b

● ● ●

● ●

●

● ●

●

●

H o t S t a r t P C R M a s t e r M i x

U p t o 3 k b

● ● ●

● ●

●

● ●

● ●

●

E f fi - T a q ™ D N A P o l y m e r a s e ( H o t S t

a r t )

U p t o 2 k b

● ● ●

●

● ●

●

● ●

●

●

P R P o l y m e r a s e

U p t o 5 k b

●

●

● ● ●

—

●

●

X T - 5 P o l y m e r a s e

U p t o 5 k b

● ● ●

●

● ● ●

●

●

●

X T - 2 0 P o l y m e r a s e

U p t o 2 0 k b

● ● ●

●

● ●

●

●

●

●

G o o d

● ●

B e t t e r

● ● ●

B e s t

●

T o b e s u p p l e m e n t e d w i t h

M A G I C A m p l i fi c a t i o n S o l u t i o n


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Ordering Information :THERMOPHILIC POLYMERASES

Taq DNA Polymerase (Supplied with 10X Buffer containing Gelatin and 15 mM MgCl₂)

Cat No. PI. No. Product Unit Price `

610601600031730 MME5L Taq DNA Polymerase (3 U/µl) 250 U (Includes Enzyme: 1 vial; 10X Taq Buffer A: 1 vial) 250 U 2290

610601600051730 MME5J Taq DNA Polymerase (3 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer A: 4 vials) 1000 U 4350

610601600061730 MME5B Taq DNA Polymerase, 2 x 2500 U (3 U/µl) ( Includes Enzyme: 2 vial ; 10X Taq Buffer A:

7 bottles x 3 ml), 5000 U

21000

610602300051730 MME23L Taq DNA Polymerase (5 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer A: 4 vials) 1000 U 4350

610602300061730 MME23B Taq DNA Polymerase, 2 x 2500 U (5 U/µl) (Includes Enzyme: 2 vials; 10X Taq Buffer A:

7 bottles x 3 ml), 5000 U

21000

610602400051730 MME24L Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer A: 4 vials) 1000 U 4350

Taq DNA Polymerase(Supplied with 10X Buffer containing Gelatin, separate vial of 25 mM MgCl₂)

610602500051730 MME25J Taq DNA Polymerase (3 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer B: 4 vials; 25 mM MgCl₂:

4 vials) 1000 U

4350

610602500051730 MME25B Taq DNA Polymerase, 2 x 2500 U (3 U/µl) (Includes Enzyme: 2 vials; 10X Taq Buffer B:

7 bottles x 3 ml; 25 mM MgCl2: 7 bottles x 3 ml), 5000 U

4250

610602700051730 MME27L Taq DNA Polymerase (5 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer B: 4 vial; 25 mM MgCl₂:

4 vials) 1000 U

4350

610602700061730 MME27B Taq DNA Polymerase, 1 x 1000 U (5 U/µl) (Includes Enzyme: 4 vials; 10X Taq Buffer B:

14 bottles x 3 ml; 14 bottles of 25 mM MgCl2), 50000 U

21000

610602800051730 MME28L Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer B: 4 vials; 25 mM MgCl₂:

4 vials) 1000 U

4350

Taq DNA Polymerase(Supplied with 10X Buffer containing TritonX-100 and 15 mM MgCl₂)

Cat No. PI. No. Product Unit Price

`

610602900051730 MME29J Taq DNA Polymerase (3 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer E: 4 vials) 1000 U 4250

610603000051730 MME30L Taq DNA Polymerase (5 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer E: 4 vials) 1000 U 4250

610603100051730 MME31L Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer E: 4 vials) 1000 U 4250

Taq DNA Polymerase(Supplied with 10X Buffer containing TritonX-100, separate vial of 25 mM MgCl₂)

610603200051730 MME32J Taq DNA Polymerase (3 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer F: 4 vials; 25 mM MgCl₂:

4 vials) 1000 U

4250

610603300051730 MME33L Taq DNA Polymerase, 4 x 2500 U (5 U/µl) (Includes Enzyme: 4 vials; 10X Taq Buffer F:

14 bottles x 3 ml:: 14 bottles of 25 mM MgCl2), 10000 U

4250

610603400051730 MME34L Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer F: 4 vials; 25 mM MgCl₂:

4 vials) 1000 U

4250

Red Taq DNA Polymerase

(Supplied with 10X Buffer containing Gelatin and 15 mM MgCl₂)Cat No. PI. No. Product Unit Price

`

610603500051730 MME35J Red Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buf fer A: 4 vials) 1000 U 6240

Red Taq DNA Polymerase(Supplied with 10X Buffer containing Gelatin and a separate vial of 25 mM MgCl₂)

610603600051730 MME36J Red Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buffer B: 4 vials;

25 mM MgCl₂: 4 vials) 1000 U6480

Red Taq DNA Polymerase(Supplied with 10X Buffer containing TritonX-100 and 15 mM MgCl₂)

610603700051730 MME37J Red Taq DNA Polymerase (1 U/µl) (Includes Enzyme: 1 vial; 10X Taq Buf fer E: 4 vials) 1000 U 8930


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PCR Master Mix (2X)PCR Master Mix is Ready To Use reagent formulation in 2Xconcentration ideal for instant and efficient amplification. TheMaster mix combines Genei's Taq DNA Polymerase, dNTPs and

an optimum reaction buffer, This ready to use mix providesrobust and reliable performance for routine PCR applications.

Storage : -20°C

Highlights :

• Easy to use.• Reduced experimental variabilities.• Minimal risk of contamination.• Consistent results.• Saves reaction setup time.

Ordering Information :Cat # PI. # Product Unit Price

`

610602200031730 MME22 Master Mix (2X) (without

100 bp DNA Ladder) (5 x

100 reactions)

10500

610602200051730 MME22L Master Mix (2X) (without

100 bp DNA Ladder) (20 x

100 reactions)

31500

PCR Master Mix Kit (2X)This kit consists of all the reagents required to perform 50standard PCR reactions.

Storage : -20°C


`

610667700041730 KT77 PCR Master Mix Kit (2X)

(with 100 bp DNA Lad-

der) (100 reactions)

6200

DNA Amplification Reagent Kit (withmarker) 100 reactions

DNA Amplification Kit contains all essential reagents packed for50/100 individual reactions. The reagents supplied are of highestquality and are guaranteed to perform DNA amplification.

Control DNA included with suitable primers to perform 20 tests.Application :

• Polymerase chain reaction or PCR is an effective tool tosynthesise large quantities of defined sequences of DNA usingTaq DNA Polymerase, dNTPs, two oligonucleotide primersthat hybridize to opposite strands and flank the target DNAsequence to be amplified.

• Using this kit one can amplify target DNA of varied sizesranging from 100 bp to 3 kb.

Storage : -20°C

Taq DNA PolymeraseTaq DNA Polymerase is a 94 KD thermostable enzyme. Its optimumtemperature of activity is between 55° C and 75° C. The enzyme lacks3' to 5' exonuclease (proof reading) activity but has an inherent

5' to 3' exonuclease activity.Specification : The enzyme is supplied at concentrations of 1 U/µl,3 U/µl, 5 U/µl.

Taq DNA Polymerase Buffers : Refer the chart for buffercomposition.


Storage and Dilution buffer :

Tris-HCl (pH 8), KCl, EDTA, DTT, Tween 20, Igepal and Glycerol.

Storage : -20°C

Application :

Amplification of DNA fragments of varied sizes ranging fromabout 100 bp to 3kb by polymerase chain reaction.

The enzyme can be used in RAPD studies to detect polymorphismin several species.

Taq DNA Polymerase can be supplied with :

10X buffer containing 15 mM MgCl₂ or

10X buffer and separate vial of 25 mM MgCl₂

Note : Concentration of the components of the buffer areproprietary.

Taq DNA Polymerase BuffersBuffer Conc. 10X Buffer

Composition

Taq Buffer A 10X Tris (pH9.0)

KCl

15 mM MgCl₂

Gelatin

Taq Buffer B 10X Tris (pH9.0)

KCl

Gelatin

Taq Buffer E 10X Tris (pH 9.0)

KCl

15 mM MgCl₂

TritonX-100

Taq Buffer F 10X Tris (pH 9.0)

KClTritonX-100

Storage : -20°C



`

610653100011730 METB1S Taq Buffer A 1 ml 1160

610653200011730 METB2S Taq Buffer B 1 ml 1160

610653300011730 METB13 Taq Buffer E 1 ml 1200

610653400011730 METB14 Taq Buffer F 1 ml 1270


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`

610660300051730 KT03 DNA Amplification

Reagent Kit (with marker)

100 reactions

13910

610660500011730 KT03I DNA Amplification Kit

(with marker) 50 reactions

7980

DNA Amplification Core Kit (withmarker) 100 reactions

DNA Amplification Core kit contains all essential reagentspacked for 100 individual reactions. This kit does not havethe control DNA and primers which are supplied with DNAAmplification Reagent kit.

Cat # 610660400051730 Control DNA and Primers are notsupplied with this kit.

Storage : -20°C


`

610660400051730 KT03C DNA Amplification Core

Kit (with marker) 100

reactions

10500

Red Dye PCR Master Mix (2X)Red Dye PCR Master Mix is Ready To Use reagent formulation in2X concentration ideal for instant and efficient amplification.

The Red Dye PCR Master Mix combines Genei's Red Taq DNAPolymerase(consist of unique inert red dye), dNTPs and anoptimum reaction buffer, After amplification, the product canbe directly loaded onto agarose gel without the addition of gelloading dye. This ready to use mix provides robust and reliableperformance for routine PCR applications.

Storage : -20°C

Highlights :

• Easy to use.• Product can be directly loaded on to gel.• Reduced experimental variabilities.•

Minimal risk of contamination.• Consistent results.• Saves reaction setup time.



`

610602100031730 MME21 Red Dye PCR Master Mix

(2X) (5 x 100 reactions)

10400

610602100051730 MME21L Red Dye PCR Master

Mix (2X) (2000 reactions)

32700

Red Dye PCR Master Mix Kit (2X)This kit consists of all the reagents required to perform 50standard PCR reactions.

Storage : -20°C

Materials Provided :

• 5 vials of Red Dye PCR Master Mix(2X)• Sterile water• StepUp™ 100bp DNA ladder (Ready to use)



`

610667800041730 KT78 Red Dye PCR Master Mix

Kit (5 x 20 reactions)

6320

HotStart Taq DNA PolymeraseMost thermophilic polymerases exhibit significant polymeraseactivity even at ambient temperatures resulting in lowsensitivity and non-specificity leading to misinterpretation ofresults specifically in clinical applications. This can be avoidedby adding enzyme after the initial denaturation (Manual HotStart). Hence there has been a clear trend towards the useof Thermostable Polymerases that remain inactive at lowertemperatures (below 50°C).

HotStart Taq DNA Polymerase is an optimized mixture ofhighly purified Taq DNA Polymerase with high affinity anti-Taqmonoclonal antibody that inhibits polymerase activity by

binding to Taq DNA Polymerase. The enzyme remains inactiveuntil the reaction mixture reaches higher temperature. Completeactivation is restored after the initial denaturation step at 94°C for2-5 minutes, thereby providing an automatic "Hot start" forTaq DNA Polymerase.

Performance Test:

• Hotstart Taq DNA Polymerase is tested extensively for itsreproducible performance in critical PCR amplifications andin Reverse Transcription-PCR.

• Enzyme is tested for amplification of 131 bp fragment of TNFgene and 400 bp fragment of ß−globin gene from humangenomic DNA using non-optimal primers.

• Enzyme is tested for detecting different subtypes of HumanPapilloma virus (HPV) using consensus primers.

Storage : -20°C


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Ordering Information :HotStart Taq DNA Polymerase

(Supplied with 10X Buffer containing Gelatin and 15 mM MgCl₂)


`

610602000031730 MME20L HotStart Taq DNA Polymerase (3 U/µl) 250 U (Includes Enzyme:

1 vial; 10X Taq Buffer A: 1 vial)

6300

610602000051730 MME20J HotStart Taq DNA Polymerase (3 U/µl) 1000 U (Includes Enzyme:

1 vial; 10X Taq Buffer A: 4 vials)

24260

HotStart Taq DNA Polymerase(Supplied with 10X Buffer containing Gelatin and a separate vial of 25 mM MgCl₂)

610604000051730 MME40J HotStart Taq DNA Polymerase (3 U/µl) 1000 U (Includes Enzyme:1 vial; 10X Taq Buffer B: 4 vials; 25 mM MgCl₂: 4 vials)

24260

HotStart PCR Master Mix (2X)HotStart PCR Master Mix (2X) is ready to use reagent formulationat 2X concentration for performing Hot Start PCR. The Mastermix combines HotStart Taq DNA Polymerase, nucleotides and anoptimum reaction buffer. This ready to use mix provides robustand reliable performance for demanding PCR applications inwhich high specificity and sensitivity are desired.

Performance Test :

HotStart PCR Master Mix (2X) is tested extensively for itsreproducible performance in critical PCR amplifications.

Enzyme is tested for amplification of 131 bp fragment of TNFgene and 400 bp fragment of ß−globin gene from humangenomic DNA using non-optimal primers.

Enzyme is tested for detecting different subtypes of HumanPapilloma virus (HPV) using consensus primers.

Application :

• Enhancement of specificity and sensitivity in the detectionof low copy number templates in complex DNA background.

• For Use in diagnostic labs to detect both infectious and

genetic disorders by DNA based diagnosis.• Variations in setup time does not affect the final product in

Highthroughput PCR.

Highlights :

• Highly specific• Low background• Higher yields• Easy to use• Saves time and effort• Variation in set up time do not affect reproducibility• Overcomes limitation of manual hot start

HotStart Taq DNA Polymerase Assay buffer (1X) :

Tris-HCl (pH 9.0), KCl, 1.5mM MgCl₂ and Gelatin.

Storage and Dilution buffer :

Tris-HCl (pH 8.0), KCl, EDTA, DTT, Tween 20, Igepal and Glycerol.

Specification : The enzyme is supplied at a concentration of 3U/µl and 1 U/µl.

With every order of HotStart Taq DNA Polymerase we supply

(a) 10X Assay buffer(b) 25 mM MgCl₂

Note : Concentration components of the buffer are proprietary.

Highlights :

• Hot Start format.• Higher sensitivity and specificity.• Reduced experimental variabilities.• Ideal for high throughput PCR.• Minimal risk of contamination.• Highly consistent results.• Room temperature reaction assembly.•

Saves reaction setup time.


`

610604200011730 MME42S HotStart PCR Master Mix

(2X) (1 X 50 reactions)

5290

610604200031730 MME42L HotStart PCR Master Mix

(2X) (5 X 50 reactions)

17990


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PR PolymerasePR Polymerase is a thermostable, DNA-dependent, DNApolymerase that exhibits robust Proof Reading activity. Theenzyme exhibits optimum polymerization activity at 72°C

in presence of Mg²+ions. The 3’ to 5’ exonuclease mediatedproof reading activity enables the enzyme to proofreadmis-incorporated nucleotides and reduce errors duringpolymerization.

PR Polymerase is the enzyme of choice for high fidelity DNAsynthesis. It can be efficiently utilized for high fidelity PCRamplification of DNA fragments to be used for gene cloning,site-directed mutagenesis and other genetic manipulations.

Assay Buffer (1X): Tris-HCl (pH 8.8), (NH₄)₂ SO₄, KCl, 2mMMgSO₄, Triton X 100 and Nuclease free BSA.

Specification :

Enzyme is supplied at a concentration of 3 U/µl. With everyorder of PR Polymerase we supply:

PR Polymearse 3U/µl

10X PR polymerase assay buffer: 2 vials

PR Polymerase is recommended for use in PCR and primerextension reaction that require high fidelity. The product canbe efficiently used for :

• Gene cloning• DNA expression• Mutation analysis and

• Other genetic manipulations.Note: Concentration of the components of the buffer areproprietary.

Performance Test :

PR Polymerase is tested extensively for PCR amplifications of specificDNA fragments using human genomic DNA andλ DNA as templates.The conditions need to be optimized for each individual system.It is observed that for products 1.5kb, optimization is required.

Storage : -20°C



`

610601700031730 MME17M PR Polymerase, 250 U,

(Enzyme 3U/µl: 1 vial;

10X PR Polymerase Assay

Buffer: 1 vial)

8400

XT-5 PCR SystemXT-5 PCR System is useful to amplify upto 5 kb target DNA fromgenomic DNA templates. A well-defined ratio of enzymes alongwith a set of highly optimized buffer systems (Assay buffers 5A

and 5B) ensures specific PCR products. Assay buffer 5A is usedfor obtaining highly specific target DNA with suboptimal yieldwhile assay buffer 5B is best utilized for higher yields.

XT-Polymerase buffer 5A : (1X)

TAPS (pH 8.8), KCl, 1.75mM MgCl₂ and Gelatin.

XT-Polymerase buffer 5B : (1X)

Tris (pH 9.1), (NH₄)2SO₄, 1.5mM MgCl₂.

Buffers supplied at 10X concentration.

Specification :

Enzyme is supplied at a concentration of 3U/µl. With every orderof XT-5 PCR system we supply:

a. Enzyme: 1 vialb. 10X assay buffer 5A: 1 vialc. 10X assay buffer 5B: 1 vial

Application :

• Useful for high efficiency PCR amplifications. The productsobtained can be used for gene cloning and other geneticmanipulations.

• Ensures higher yields of the amplified product with improvedfidelity.

Note: Concentration of the components of the buffer areproprietary.

Performance Test :

XT-5 PCR system is tested for PCR amplifications of fragmentsizes - 1.5 kb to 8.0 kb using both human genomic DNA andlambda DNA as templates. Sequence specific primers were usedfor amplifications.

Storage : -20°C



`

610601800031730 MME18L XT-5 PCR system 250 U,

(Enzyme: 1 vial; 10X Assay

Buffer 5A: 1 vial; 10X As-

say Buffer 5B: 1 vial) 1 EA

3680

610601800051730 MME18J XT-5 PCR system 1000 U,


Buffer 5A: 2 vial; 10X As-

say Buffer 5B: 2 vial)

11550


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XT-20 PCR SystemXT-20 PCR system is useful in amplifying long target DNA upto20 kb from genomic DNA templates. A well-defined ratio ofenzymes along with a set of highly optimized buffer systems

(Assay buffers 20A and 20B) ensures specific PCR products.Assay buffer 20A is optimized for amplifications of 8.0 kb to12.0 kb target sequences and Assay buffer 20B is optimized for12.0 kb to 20.0kb amplifications.

XT-Polymerase buffer 20A : (1X)

Tris (pH 9.1), (NH4)2SO4, 2 mM MgCl₂.

XT-Polymerase buffer 20B : (1X)

Tris (pH 9.1), (NH4)2SO4, 2.75 mM MgCl₂.

Buffers supplied at 10X concentration.

Specification :

Enzyme is supplied at a concentration of 3 U/µl. With everyorder of XT-20 PCR system :

For MME19S and MME19L (Enzyme : 1 vial, 10X assay buffer20A: 1 vial, 10X assay buffer 20B: 1 vial)

For MME19J (Enzyme : 1 vial, 10X assay buffer: 2 vials, 10Xassay buffer 20B: 2 vials)

Application :

XT-20 PCR system is useful for amplifying long targets ofgenomic DNA up to 20.0kb.

• Can be successfully used for mapping and sequencing

genomes and for analyzing eukaryotic genes.• Can be used in diagnosis of human disease genes that often

exceed a length of 10.0 kb.Note : Concentration of the components of the buffer areproprietary.

Performance Test :

XT-20 PCR system is tested for PCR amplifications of fragmentsizes - 8.0 kb to 25.0 kb, using both human genomic DNA andlambda DNA as templates. Sequence specific primers were usedfor amplifications.

Storage : -20°C



`

610601900031730 MME19L XT-20 PCR system 250 U,


Buffer 20A: 1 vial; 10X

Assay Buffer 20B: 1 vial)

3820

610601900051730 MME19J XT-20 PCR system 1000 U,


Buffer 20A: 1 vial; 10X

Assay Buffer 20B: 2 vial)

12100

Long PCR 20kb Ampli KitLong PCR 20 kb Ampli kit is ideal for amplifications of longertarget DNA fragments, upto 20kb, from genomic DNA templatesusing sequence specific primers. The kit provides all essential

reagents to perform 50 individual reactions. Control DNA andsequence specific primers are included to perform 10 controlreactions.

• XT-20 PCR system is a unique enzyme mix that combines thepowerful 5'-3' polymerase activity of Taq DNA polymeraseand the 3'-5' exonuclease mediated proofreading activity ofPR polymerase, resulting in high fidelity PCR products.

• Highly optimised buffer systems (XT-Polymerase buffers 20Aand 20B) guarantee better performance.

• Human genomic DNA with sequence specific primers toamplify 15.0 kb DNA fragment of tPA gene for 10 controlamplification reactions are provided.

Application :

• Useful for mapping and sequencing genomes and analyzingeukaryotic genes.

• Can be used in diagnosis of human disease genes that oftenexceed a length of 10.0 kb.

Storage : -20°C


`

610667600011730 KT76 Long PCR 20 kb Ampli Kit

(50 reactions)

9240

RAPD Primer SetsRAPD (Random Amplified Polymorphic DNA) is a polymorphismassay which is based on the amplification of random DNAsegments using sets of primers of arbitrary nucleotide sequence.In theory, the primer anneals to many regions of the genomesimultaneously. However, geometric amplification only occursin those regions in which the 3' end of the annealed primersface one another on opposite strands and are no more than3 Kb apart. Thus, the technique essentially scans a genome forthese small inverted repeats and amplifies intervening DNAsequences of variable length.

The RAPD Primer Set is designed for different organisms. Thereare five different primer sets available for bacteria, fungi,plants, animals and humans. It enables the user to choose theappropriate primer set for fingerprinting.


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Cat No. PI. No. Product Primers(no) O.D (µg) Unit Price

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610692100251730 RBa-D Bacterial Primer Set - Full 25 99 35840

610692300251730 RFu-D Fungal Primer Set - Full 25 99 35840

610692800251730 RPi-D Plant Primer Set - Full 25 99 35840

AFLP Primer Set for Small and LargeGenomesThe Amplified Fragment Length Polymorphism (AFLP) techniqueis one of a number of DNA fingerprinting procedures that takesadvantage of the polymerase chain reaction (PCR) to amplify alimited set of DNA fragments from a specific DNA sample (Voset al. 1995; Blears et al 1998). AFLP's are quickly becoming the

tool of choice for many applications and organisms. Potentialapplications include screening DNA markers linked to genetictraits, parentage analysis, forensic genotyping, diagnosticmarkers for pathogen borne diseases, and population genetics.Since the AFLP technique can be applied to a wide variety oforganisms with no prior sequence information this techniquehas the potential to become a universal DNA fingerprinting tool.

Product information: The AFLP primer set for small and largegenomes provides sufficient primers for 100 reactions.

Highlights :

• Adapters, Pre-selective and selective primers supplied for

conducting AFLP experiment.• Does not include other PCR consumables which are readily

available.• Excellent resolution obtained as checked against wide range

of organisms with small genome sizes like various bacteria(both gram positive and gram negative), fungi, yeast etc.

Storage : -20°C

Ordering Information:


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610694200011730 APS100 AFLP Primer Set for

Small Genomes

58650

610694100011730 APL100 AFLP Primer Set for

Large Genomes

61410

PCR KlenZol™PCR KlenZol™ provides a single step, fast and efficient methodfor purification of amplified PCR products post amplification.This is a simple and effective means of removing thecontaminants in an amplified product which in turn hamper thedownstream processing of the amplified product. It is solutionbased and does not involve any binding to membranes/beads,

thus reducing the chances of loss of amplified product duringthe purification step. PCR KlenZol™ uses specialized additiveswhich selectively adsorb DNA thus removing the leftover primer,primer dimer, dNTPs, salts, enzyme and mineral oil.

Product Description : PCR KlenZol™ (2X) provides 5 ml ofsolution.

Recommended Use : 1X

Storage : Store the solution at Room TemperatureHighlights:

• Single step, effective and simple protocol for clean up ofamplified product.

• 80 to 90% recovery of amplified product.• Only solution based and does not involve columns or beads,

which makes it easy to scale up.• Volume of solution to be used is equal to the amount of

amplified product.• Purified product is ready for down processing like cloning,

restriction analysis, sequencing.• Can be used for product volume of 10 - 100 µl.• Primer-dimer removal - effectively removes primer-dimer

(>90%) for PCR product size ranging between 300-5000 bp.• Effectively removes contaminants from amplified products

ranging in size from 100-5000 bp.



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612107180051730 FC71 PCR KlenZol™ 5*1ml),

5ml

15900

dNTP Solutions (Adjusted to Neutral pH)Deoxynucleoside triphosphates are supplied as sodium saltsolutions.

Application : Routinely used for

• DNA sequencing• Labelling and• PCR techniques

All dNTP solutions supplied in 10 mM or 100 mM concentration.

Storage : -20°C


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dNTP Mix

610652300031730 FC23J dNTP Mix, 10 mM(2.5 mM each) (1000 µl)

4680

610652400021730 FC24L dNTP Mix, 10 mM

(2.5 mM each) (4 x 200 µl)

3430

610652400041730 FC24J dNTP Mix, 10 mM (2.5 mM

each) (4 x 1000 µl)

11440

610652200011730 FC23HL dNTP Mix, 100 mM

(25 mM each) (100 µl)

3890

dNTP Set610651200011730 FC10 dNTP Set, 10 mM (4 x 100 µl) 2100

610651200031730 FC10J dNTP Set, 10 mM (4 x 500 µl) 5250

610651300021730 FC13L dNTP Set, 10 mM

[5 (4 x200 µl)] (20 vials)

11600

610651000031730 FC10HJ dNTP Set, 100 mM

(4 x 250 µl)

9180

dNTP Solutions610650600011730 FC6HL dATP, 100 mM (100 µl) 2940

610650700011730 FC7HL dGTP, 100 mM (100 µl) 2940

610650800011730 FC8HL dTTP, 100 mM (100 µl) 2940

610650900011730 FC9HL dCTP, 100 mM (100 µl) 2940

Mineral OilLight White Oil

Used in high temperature reactions to protect sample fromevaporation eg. PCR reactions.

Does not contain stabilizer.Storage : Room temperature.


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613601281001730 FC21L Mineral Oil 100 ml 5090

DNA Diagnosis & ReagentsWhite Spot Syndrome Virus (WSSV)Detection Kit (Nested PCR method)

White Spot Syndrome Virus (WSSV) has caused mass mortalitiesand devastating production losses to shrimp farming in manycountries. It is now evident that infected seed is a commonsource of WSSV infection in shrimp ponds and that PCRscreening of seed can significantly reduce the risk of crop failure.

Conventional diagnosis fails to detect early stages of infection.WSSV detection kit, based on Nested PCR method is a sensitiveand specific diagnostic tool that can detect infection even ata very early stage.

Highlights :

• Follows nested PCR principle - High sensitivity and specificityof virus detection.

• Detects up to 2 viral particles.•

Fast and simple DNA extraction protocol.• Convenient aliquots of reagents - Reduced chances of

contamination.• Sterile disposable grinders for sample processing.• The entire procedure can be completed within 4 hours.

Ordering Information :Cat No. PI. No. Product Unit Price

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610680400011730 WSSV25 WSSV Detection Kit (for

25 Tests)

13050

610680500051730 WSSV100A WSSV Detection Kit (for

100 Tests) with Gel Elec-

trophoresis consumables

37070

Single Tube WSSV Detection Kit(Single Tube Nested PCR)

Single Tube Nested PCR Kit for White Spot Syndrome Virus(WSSV) detection is based on innovative single tube nested PCRtechnique. In this assay both the first and the nested PCR stepsare performed within one PCR run in the same reaction vial.

The assay can indicate the degree of severity of infection. Thekit can produce up to three PCR products depending on theseverity of infection as below :

• Severe Infection (> 10⁵ viral particles) - produces all 3

fragments (942, 525, & 204 base pairs (bp)• Moderate Infection (10³ - 105 viral particles)- produces

2 fragments (525 & 204 bp)• Low Infection (10-10³ viral particles) - produces 1 fragment

(204 bp)

Highlights :

• A single tube nested PCR reaction - Less chances ofcontamination

• Follows nested PCR principle - High sensitivity and specificityof viral detection

• Distinguishes the degree of severity of infection Severe,Moderate and Low

• Detects up to 10 viral particles in 3.5 to 4 hours• Fast and simple DNA extraction protocol• Convenient aliquots of reagents - Ease of work and reduced

chances of contamination• Sterile disposable grinders for easy sample processing



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610680300011730 STWSSV50 Single Tube WSSV Detec-

tion Kit (50 Tests)

19800


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Amplification Reagents Set(for Mycobacterium tuberculosis )

Tuberculosis continues to be the leading cause of morbidity

and mortality by an infectious disease worldwide. Effectivetreatment of TB requires the rapid detection of Mycobacteriumtuberculosis.

This set is based on the principle of single tube nested PCRtargeting the IS6110 sequence that is present in multiple copiesin the genome of the Mycobacterium tuberculosis complex.Studies have shown that highest sensitivity is obtained byusing IS6110 nested PCR when compared to PCR targetingother regions.

The diagnosis of TB is made by microscopy and culture.Direct Microscopy lacks sensitivity and specificity. Culture ismore sensitive and is still the yardstick for diagnosis, but the

time required and frequent negative results in paucibacillaryspecimens are the limitations. PCR is widely used for thedetection of Mycobacterium tuberculosis in clinical specimens,because the tests are rapid, sensitive, and specific.

Highlights :

• Detects specifically Mycobacterium tuberculosis complexpresent in samples.

• Based on the principles of nested PCR for increased specificityand sensitivity.

• Uses HotStart Taq DNA Polymerase for increased specificity.• Carryover contamination prevention by dUTP-UDG system.• Rapid Spin column based DNA extraction for effective removal

of PCR inhibitors from the sample.• Results obtained in 4 hours.• Convenient aliquots of reagents to reduce the chances of

contamination.• Internal control DNA provided for the identification of

processed specimens containing substances that couldinhibit PCR.

• Simulated positive controls included in the set.

Note : For Research use only. Not for diagnostic applications.



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610670300011730 MTB25 Amplification Reagents Set for

Mycobacterium tuberculosis

(for 25 tests)

19320

610670300021730 MTB50 Amplification Reagents Set for

Mycobacterium tuberculosis

(for 50 tests)

34650

Amplification Reagents Set forMalarial Parasites(Plasmodium falciparum and Plasmodium vivax )

Malaria is one of the main health problems worldwide. It isestimated that it affects around 500 million people, of which2.5 million die per year. Early detection, as well as speciesidentification, is a must for an accurate treatment.

The standard method for detecting plasmodium infection isthe microscopic examination of Giemsa-stained thick bloodsmear. Although this method is effective and inexpensive, butits laborious, time consuming and sensitivity is very less.

Recently, alternative diagnostic method, such as PCR, have beenused for the detection and identification of malarial parasites.This assay detects parasites in mixed and low level infection,

being more sensitive than conventional methods.Amplification Reagent Set is based on the principle of nestedPCR targeting the 18S small subunit ribosomal RNA (ssrRNA)gene and is used for the identification of Plasmodium falciparumand Plasmodium vivax species.

Highlights :

• Rapid spin Column based DNA extraction for effective removalof PCR inhibitors from the DNA sample.

• Based on the principle of Nested PCR for increased specificityand sensitivity.

• Results obtained in 5 hrs.•

Convenient aliquots of reagents to reduce the chances ofcontamination.• Simulated positive controls included in the set.




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610670200011730 MAL25 Amplification Reagents Set

for Malarial Parasites,

25 tests

21160


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Amplification Reagents Set (for HumanPapilloma Virus)

Cancer of the uterine cervix is the most common malignanttumor in women worldwide. Human Papilloma Virus (HPV) has

been identified as the 'necessary cause' of cervical cancer. It hasbeen shown from several studies that HPV infection is a goodmarker for women with cervical neoplasia and precancerouslesion. Women persistently infected with certain "oncogenic"HPV show a high rate of progression of dysplasia to invasivecancer of the cervix. Therefore, in best traditions of medicalpractice, the clinicians are obligated to seek out the presence ofcausal agents of the disease, the oncogenic types of HPV. Thus,diagnosis of HPV infection may facilitate early identification ofwomen at increased risk of developing cervical cancer.

This set is based on the amplification of E6 and E7 genes ofoncogenic HPVs (HPV-16, 18, 31, 33, 35, 45, 52b and 58) usingconsensus primers. The E6 and E7 genes are preferentiallyconserved even when the viral DNA gets integrated into thehuman chromosome. Thus, the amplification targeting E6 andE7 region is less likely to produce false negative results.


Highlights :

• Detects oncogenic HPV types (E6 & E7) present in samples.• Based on the principle of Touchdown PCR for increased

specificity and sensitivity.• Uses HotStart Taq DNA Polymerase for increased specificity.• Rapid spin Column based DNA extraction for effective removal

of PCR inhibitors from the DNA sample.• Results obtained in 5 hrs.• Convenient aliquots of reagents to reduce the chances of

contamination.• Simulated positive controls included in the set.



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610670100011730 HPV25 Amplification Reagents Set

for Human Papilloma Virus

(25 tests)

14300

AMV Reverse TranscriptaseAMV Reverse Transcriptase is a RNA - dependent DNApolymerase purified from Avian Myeloblastosis virus. Theenzyme is used extensively in cDNA synthesis and dideoxy DNA

sequencing.The enzyme possesses several enzymatic activities whichinclude an RNA directed DNA synthesis, a DNA-dependent DNApolymerase, an RNase H activity and an unwinding activity.

Unit definition : One unit of AMV reverse transcriptase isdefined as amount of enzyme which incorporates 1 nanomoleof (3H) dTMP into an acid insoluble product in 10 minutes at37°C using poly (A) - oligo (dT) as template - primer.

Assay Buffer : (1X) 50 mM Tris-HCl (pH 8.5), 8 mM MgCl₂,30 mm KCl and 1mM DTT.

Buffer supplied as 5X concentration.

Storage buffer: 200 mM Phosphate Buffer (pH 7.2), 2 mM DTT,

0.2% Triton X-100 and 50% glycerol.Application : The enzyme is used extensively in cDNA Synthesis.

Performance Test :

Enzyme Tested for its performance in RT-PCR reactions.

Storage : -20°C


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610600900021730 MME9L AMV Reverse Transcriptase 100 4750

M-MuLV Reverse TranscriptaseM-MuLV RT is a RNA dependent DNA polymerase that usessingle stranded RNA or DNA as template. It lacks endonucleaseactivity and has lower RNase H activity.

Unit definition : One unit of M-MuLV reverse transcriptase isdefined as amount of enzyme which incorporates 1 nanomoleof (3H) dTMP into an acid insoluble product in 10 minutes at37°C using poly (A) - oligo (dT) as template - primer.

Assay Buffer : (1X) 50mM Tris-HCl (pH 8.5), 8 mM MgCl₂, 30mµ KCl and 1mM DTT.

Buffer supplied in 5X concentration.

Storage Buffer : 20 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mMEDTA, 1 mM DTT, 0.1% Igepal and 50% glycerol.

Application : The enzyme is used in cDNA synthesis and dideoxyDNA sequencing.

Performance Test :

Enzyme Tested for its performance in RT-PCR reactions.

Storage : -20°C



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610601300051730 ME13S M-MuLV RT 1000 U 4730

610601300061730 ME13L M-MuLV RT 5000 U 7520


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AMV RT-PCR KitThe amplification of RNA requires the conversion of RNAsubstrate to cDNA. This is achieved using AMV ReverseTranscriptase. The cDNA thus generated acts as the template

for subsequent PCR amplification.Two primers are provided for cDNA synthesis :

1. Random Hexamer that binds at non-specific sites alongthe mRNA template.

2. Oligo (dT)₁₈ primer that binds at the 3'-end of the poly (A)tail of mRNA.

The kit has all components necessary to generate first strandcDNA and subsequent PCR amplification. Enough materialssupplied to perform 20 reactions. We have also included controlRNA and suitable primers for 4 control experiments.

Storage : -20°C

Application :

•

RT-PCR Kit can be effectively used to detect the presenceof RNA viruses.• PCR products generated using this kit can be cloned using

standard protocols.


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610662400011730 KT24 RT-PCR Kit (20 reactions) 16450

610662400021730 KT24M RT-PCR Kit (50 reactions) 27500

M-MuLV RT-PCR KitM-MuLV RT-PCR Kit uses M-MuLV Reverse Transcriptaseenzyme for generating cDNA from RNA substrate. The cDNAthus generated acts as the template for subsequent PCRamplification.

Two primers are provided for cDNA synthesis:

Random Hexamer that binds at non-specific sites along themRNA template.Oligo (dT)₁₈ primer that binds at the 3'-end of the poly (A) tailof mRNA.

The kit has all components necessary to generate first strandcDNA and subsequent PCR amplification. Enough materialssupplied to perform 20 reactions. We have also included controlRNA and suitable primers for 4 control experiments.

Storage : -20°C

Application :

• M-MuLV RT-PCR Kit can be effectively used to detect thepresence of RNA viruses.

• PCR products generated using this kit can be cloned usingstandard protocols.

Ordering Information:Cat # PI. # Product Unit Price

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6106674000117

bioscience price list 2014-15 - volume i

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