pcr brochure, march 2002kahn_sci/flow/b2-and_f2_novagen_pcr_guide.pdfunique proofreading enzyme,...
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PCR E N Z Y M E S O V E R V I E W
2 www.novagen.com
Novagen offers a complete selection of enzymes and kitsfor PCR, featuring KOD HiFi DNA Polymerase. Thisunique proofreading enzyme, isolated from the extremethermophile Thermococcus kodakaraensis KOD1, possess-es superior processivity and fidelity that enable faster,more accurate PCR amplification than with conventionalenzymes, including Pfu DNA polymerase (1). KOD HiFiDNA Polymerase is also available in a Hot Start versionfor high specificity and increased read length (2), and as ablend (KOD XL DNA Polymerase) recommended forvery long templates (3).
NovaTaq™ DNA Polymerase is a high-purity, recombi-nant enzyme suitable for any application requiring premi-um quality Taq DNA polymerase. For increased specifici-ty and convenience with standard PCR, we also offerNovaTaq™ Hot Start DNA Polymerase and the TaqAntibody. NovaTaq Hot Start DNA Polymerase is a chem-ically modified form of Taq DNA polymerase thatbecomes active when heated at 95°C for 7–10 minutes.The Taq Antibody is available as an alternative means toprovide hot start capability to NovaTaq DNA Polymeraseas well as other sources of Taq DNA polymerase. Pleaserefer to the table above as a guide to select the appro-priate enzyme combination for your application.
PCR EN Z Y M E SE L E C T I O N GU I D E
Enzyme PCR Product Elongation Specificity Fidelity GC-rich Yield PCR ProductSize Rate Templates Ends
KOD HiFi DNA Polymerase < 6 kbp 120 bases/s ● ■ ■ blunt
KOD Hot Start DNA Polymerase < 21 kbp 120 bases/s ■ ■ ■ ■ blunt
KOD XL DNA Polymerase < 30 kbp 120 bases/s ● ■ ■ ■ Mixed (blunt and 3'-dA)
NovaTaq™ DNA Polymerase < 5 kbp 60 bases/s ● ● ● 3'-dA
NovaTaq Hot Start DNA Polymerase < 5 kbp 60 bases/s ■ ● ▲ 3'-dA
NovaTaq DNA Polymerase + Taq Antibody < 5 kbp 60 bases/s ■ ● ▲ 3'-dA
● Satisfactory ▲ Good ■ Excellent
1. Takagi, M., Nishioka, M., Kakihara, H., Kitabayashi, M., Inoue, H.,
Kawakami, B., Oka, M., and Imanaka, T. (1997) Appl. Environ. Microbiol. 63,
4504–4510.
2. Mizuguchi, H., Nakatsuji, M., Fujiwara, S., Takagi, M., and Imanaka, T.
(1999) J. Biochem. (Tokyo) 126, 762–768.
3. Nishioka, M., Mizuguchi, H., Fujiwara, S., Komatsubara, S., Kitabayashi, M.,
Uemura, H., Takagi, M., and Imanaka, T. (2001) J. Biotechnol. 88, 141–149.
3
KOD DNA Polymerase
Faster than Taq ...More accurate than Pfu...
TA B L E O F C O N T E N T S
AM P L I F Y (see page 6)
KOD POLYMERASES
■ KOD DNA Polymerase is a new, high performance DNA polymerase
KOD HiFi DNA Polymerase – Faster than Taq, more accurate than PfuKOD Hot Start DNA Polymerase – A heat activatable form of KOD HiFi DNA Polymerase
for automated PCR set-up
KOD XL DNA Polymerase – Recombinant forms of KOD Polymerase blended for long and accurate PCR
GENOMIC DNAGenomic DNA – High quality, high integrity human, animal, and yeast DNA
RT-PCR TOOLS
First Strand cDNA Synthesis Kit – Reliable preparation of templates for RT-PCR
One Step RT-PCR Kit – Convenient, one-enzyme, single buffer system for RT-PCR
TAQ POLYMERASES AND PCR KITS
■ NovaTaq™ DNA Polymerase is a premium quality recombinant form of Thermus aquaticus DNA polymerase
NovaTaq DNA Polymerase – A molecular biology lab essential
10 mM dNTP Mix
NovaTaq PCR Kit – Everything for PCR except DNA and primersNovaTaq PCR Kit PLUS – What you need for successful PCR and PCR optimizationNovaTaq PCR Master Mix – A ready-to-use 2X concentrated PCR reagent mixtureNovaTaq Hot Start DNA Polymerase – For increased specificity and convenient reaction assemblyTaq Antibody – Monoclonal antibody for automated Hot Start PCR
OR D E R I N G IN F O R M AT I O N (see page 24)
www.novagen.com4
5
CL O N E (see page 18)
CLONING KITS
Perfectly Blunt® Cloning Kits – Efficient “universal” cloning of DNA amplified by any polymerase
AccepTor™ Vector Kits – Rapid, direct cloning of DNA amplified with non-proofreading DNA polymerases
AN A LY Z E (see page 22)
DNA LADDERS AND MARKERS
Perfect DNA™ Ladders and Markers
PCR Markers
PU R I F Y (see page 14)
DNA PURIFICATION
SpinPrep™ PCR Clean-up Kit – Rapid purification of PCR products for downstream procedures
SpinPrep Gel DNA Kit – Rapid, efficient extraction of DNA from agarose gels
SpinPrep Plasmid Kit – Rapid, high quality plasmid minipreps
SpinPrep Master Kit – Combo Kit; 20 plasmid preps and 20 DNA gel extractions
PELLET PAINT® CO-PRECIPITANTS
Pellet Paint Co-Precipitant – Rapid, quantitative precipitation of DNA and RNA, including PCR clean-up
Pellet Paint NF Co-Precipitant – Non-fluorescent visible DNA co-precipitant for sequencing and other automated applications
TR A N S F O R M (see page 20)
COMPETENT CELLS
NovaBlue Singles™ Competent Cells – High efficiency transformation of E. coli in less than 8 minutes
HT96™ NovaBlue Competent Cells – High-efficiency competent cells pre-dispensed in a 96-well format
HT96 Isothermal Block – Efficient thermal transfer for uniform incubations
KOD P O LY M E R A S E S
KOD HiFi DNA Polymerase
KOD HiFi DNA Polymerase is a recombinant form of Thermococcus
kodakaraensis KOD1 DNA polymerase. It is the most efficient
thermostable DNA polymerase, exhibiting higher accuracy, elongation
rate and processivity than any other commercially available DNA poly-
merase. KOD HiFi DNA Polymerase possesses 3' 5'
exonuclease-dependent proofreading activity that enables the
polymerase to correct nucleotide misincorporation. The enzyme
generates blunt-ended PCR fragments that are suitable for cloning
with Novagen’s Perfectly Blunt® Cloning Kits.
Each order also includes 10X KOD HiFi DNA Polymerase Buffer 1 and
10X KOD HiFi DNA Polymerase Buffer 2, plus separate vials of 25 mM
MgCl2 and 2mM dNTP Mix.
ADVANTAGES
• Higher fidelity than Pfu DNA Polymerase (1)
• Higher processivity – sequential nucleotide polymerization
is between 10 and15X greater than Pfu DNA Polymerase and
Deep Vent* DNA Polymerase (1)
• Greater yield – extension speed is 2X faster than Taq DNA
polymerase and 5X faster than Pfu DNA polymerase (1)
• More accurate PCR in a shorter time
• No truncated amplification products in PCR reaction
1. Takagi, M., et al., (1997) Applied and Environmental Microbiology 63, 4504–4510.
† Fidelity was measured by the authors as mutation frequency in PCR products using a sensitive blue/white phenotypic assay with a 5.2 kbp lacZ plasmid as template.
* Deep Vent is a trademark of New England Biolabs, Inc.
Enzyme
Species
Fidelity †
Elongation rate(bases/second)
Processivity(nucleotide bases)
KOD HiFiDNA Polymerase
Thermococcuskodakaraensis
3.5 x 10–3
106–138
> 300 bases
PfuDNA Polymerase
Pyrococcusfuriosis
3.9 x 10–3
25
< 20 bases
TaqDNA Polymerase
Thermusaquaticus YT-1
1.3 x 10–2
61
not determined
A M P L I F Y
Pure recombinant high fidelity DNA polymerases from Thermococcus kodakaraensis
PCR products amplified using KOD HiFi DNA Polymerase
DNA fragments from various templates were amplified using 2.5units KOD HiFi DNA Polymerase in a standard 100 µl reaction.Portions of each amplification reaction were analyzed by agarosegel electrophoresis (1.2% TAE).
1 2 3 41 Perfect DNA™ Markers, 0.5–12 kbp
2 5.4 kbp PCR product (lambda DNA)
3 2.0 kbp PCR product (plasmid DNA)
4 1.6 kbp PCR product (human genomic DNA)
bp
12,000 –10,000 –
8000 –
6000 –
4000 –
3000 –
2000 –
1500 –
1000 –
Ultra high fidelity DNA polymerase
www.novagen.com6
←
P C R
Lambda DNA amplification
The indicated fragments oflambda DNA were amplifiedwith KOD Hot Start DNAPolymerase
M=Markers
Genomic DNA amplification
The human myosin heavy chaingene (8.4 kbp) and human ß-globingene (12.3 kbp) were amplifiedusing KOD Hot Start DNAPolymerase
M=Markers
hh ii gg hh ff ii dd ee ll ii tt yy
KOD Hot Start DNA Polymerase
KOD Hot Start DNA Polymerase is a premixed complex of KOD HiFi
DNA Polymerase and two monoclonal antibodies that inhibit the
enzyme’s DNA polymerase and 3' 5' exonuclease activities during PCR
assembly. KOD Hot Start DNA Polymerase combines the high fidelity, fast
extension speed, and outstanding processivity of KOD HiFi DNA
Polymerase with the high yield and specificity of the antibody-based Hot
Start technology. KOD Hot Start DNA Polymerase generates blunt-ended
PCR fragments that are suitable for cloning with Novagen’s Perfectly
Blunt® Cloning Kits.
Each order also includes 10X KOD Hot Start DNA Polymerase Buffer
plus separate vials of 25 mM MgSO4 and 2mM dNTP Mix.
ADVANTAGES
• Highest accuracy, yield and processivity among
commercially available proofreading DNA polymerases (1)
• Amplifies genomic DNA templates up to 12 kbp
• Amplifies plasmid DNA templates up to 21 kbp
• Successfully amplifies GC-rich sequences
• Eliminates mispriming and primer-dimer formation
• Convenient room temperature set-up
For ordering information, see page 24.
Heat-activatable form of KOD HiFi DNA Polymerase for automated PCR set-up
M 1 2 4 6 8 10 12 15 21 M (kbp)M 8.4 12.3 (kbp)
←
l o n g
KOD XL DNA Polymerase is an optimized blend of KOD HiFi DNA
Polymerase and a mutant form of KOD HiFi that is deficient in 3' 5'
exonuclease activity. The enzyme mixture is designed for reliable
amplification of long, complex targets with robust yield and high
accuracy. KOD XL DNA Polymerase generates PCR products suitable
for cloning with Novagen’s AccepTor™ Vector and Perfectly Blunt®
Cloning Kits.
Each order also includes 10X KOD XL DNA Polymerase Buffer and
2 mM dNTP Mix.
ADVANTAGES
• Ideal for amplification of large DNA fragments
from purified DNA or crude samples
• Amplifies DNA templates up to 30 kbp
• Successfully amplifies GC-rich sequences
• Efficiently incorporates derivatized dNTPs
KOD XL DNA Polymerase
KOD P O LY M E R A S E S
Genomic DNA
Novagen’s genomic DNAs are qualified for genomic analysis including
PCR and library construction. At least 90% of the DNA is greater than
100 kbp in size, as analyzed by CHEF gel electrophoresis (> 50 kbp for
Saccharomyces cerevisiae). No inhibition of restriction enzyme activity is
observed, and the DNA is free of contaminating RNA and protein.
A M P L I F Y
ORGANISM SOURCE
Bovine blood
Cat blood
Chicken blood
Dog blood
Human (female only) blood
Human (male only) blood
Human (male & female) blood
Mouse blood
Pig blood
Rat (Sprague Dawley) blood
S. cerevisiae (wild type S288C) mixed culture
All DNAs are supplied at 100–300 µg/ml in TE buffer,100 µg per vial.
High performance enzyme blend for long and accurate PCR
High molecular weight, high purity DNA for any application
CHEF analysis
bovi
ne
pig
λla
dder
dog chicken bovine pig
M U E S U E S U E S U E S
0.7% agarose gel showingdigestion of genomicDNA (5 µg/lane, gelstained with ethidiumbromide).
U = no enzymeE = EcoR IS = Sau3A I
kbp
23 –
9.4 –6.7 –
2.3 –2.0 –
kbp
– 468
– 195
– 39
Restriction enzyme analysis
Performance comparisonλ DNA fragments were amplified using Taq DNAPolymerase and KOD XL DNA Polymerase
M λHind III DNA Markers1 1 kbp PCR product2 2 kbp PCR product3 4 kbp PCR product4 6 kbp PCR product
5 8 kbp PCR product6 10 kbp PCR product7 12 kbp PCR product8 15 kbp PCR product
8
←
A C C U R A T E
The One Step RT-PCR Kit is designed for rapid, sensitive detection of
gene expression in tissues and cells. The kit takes advantage of the
properties of recombinant Thermus thermophilus (rTth) DNA
Polymerase, which acts as both a thermostable RNA-dependent DNA
polymerase and a DNA-dependent DNA polymerase. In a single
reaction, cDNA is synthesized from input RNA through reverse
transcription, followed by PCR amplification of the cDNA without
changing the buffer or adding reagents.The amplified DNA is normal-
ly analyzed by agarose gel electrophoresis.
Use of gene specific primers or oligo(dT) and one gene specific 5'
primer is recommended with the kit. Amplification of a specific
message requires approximately two hours.
Each kit provides sufficient reagents to perform 50 RT-PCR reactions.
Positive Control RNA and Control Primers are also included.
ADVANTAGES
• Robust one-step, one-enzyme system for easy
reaction assembly
• Eliminates the risk of cross-contamination associated with two
step RT-PCR protocols
• High temperature (60°C) reverse transcription enhances
read-through of RNA secondary structure or high
GC content
• Ideal for gene expression studies
One Step RT-PCR Kit
First Strand cDNA Synthesis Kit
RT- PCR T O O L S
The First Strand cDNA Synthesis Kit is designed for the preparation
of high quality first strand cDNA from cellular RNA templates.The kit
contains MMLV Reverse Transcriptase for superior yields of full-length
cDNA. Both oligo(dT) and random hexamer primers are included for
a choice of general priming strategies as alternatives to user-supplied
specific primers. The reaction conditions are compatible with direct
addition of primers and KOD Hot Start DNA Polymerase or KOD XL
DNA Polymerase for amplification of the cDNA product.
ADVANTAGES
• MMLV Reverse Transcriptase provides superior yields of
full-length cDNA
• Both oligo(dT) and random hexamer primed synthesis options
• Kit configured for direct addition of reagents and primers
• Positive Control RNA and Control Primers included in kit
For ordering information, see page 24.
Reliable preparation of templates for RT-PCR
Convenient one-enzyme, single buffer system for RT-PCR
PCR of first strand cDNA
The Positive Control RNA was subjected tofirst strand cDNA synthesis with various primercombinations followed by addition of the 5'sense Control Primer (and antisense ControlPrimer, where appropriate) and amplification.First strand primers are indicated.AS = antisense Control Primer, RH = randomhexamers, dT = oligo(dT)
AS
AS
+ R
H
dT AS
+ d
T
RH
M
TA Q P O LY M E R A S E S A N D PCR K I T S
This dNTP Mix is a ready-to-use preparation of ultrapure dATP,
dCTP, dGTP, and dTTP (monosodium salts) at a concentration of
10 mM each in sterile deionized water at pH 7.0.The dNTP Mix is free
of RNase and DNase and is qualified for any application that
requires pure deoxynucleotides, such as PCR, cDNA synthesis, and
fill-in reactions.
10 mM dNTP Mix
NovaTaq™ DNA Polymerase is suitable for a wide range of PCR
applications. Each preparation is quality tested to ensure the highest
purity and reproducible performance. The enzyme leaves single
3'-dA overhangs that make the products suitable for cloning with
Novagen’s AccepTor™ Vector and Perfectly Blunt® Cloning Kits.
Each order also includes optimized 10X NovaTaq Buffer containing
MgCl2 for routine amplification conditions, plus separate vials of 10X
NovaTaq Buffer without MgCl2 and 25 mM MgCl2 to enable
convenient optimization of Mg2+ concentrations.
NovaTaq™ DNA Polymerase
NovaTaq PCR Kit
The NovaTaq PCR Kit includes all the reagents necessary for PCR
amplification except primers and template. Each component of the kit
has been quality tested in PCR applications. Sufficient amounts of
reagents are provided for 200 standard 50 µl (or 100 × 100 µl) ampli-
fication reactions.
The kit includes NovaTaq DNA Polymerase (250 U at 5 U/µl), 10X
NovaTaq Buffer with MgCl2, 10X NovaTaq Buffer without MgCl2, 25
mM MgCl2, 10 mM dNTP Mix and PCR Grade Water.
A M P L I F Y
A premium quality recombinant form of Thermus aquaticus DNA polymerase
PCR products amplified using NovaTaq DNA Polymerase
DNA fragments 0.5 kbp to 7.35 kbp in size were amplified using2.5 units NovaTaq DNA Polymerase in a standard 100 µl reac-tion. Products from each amplification reaction were analyzed byagarose gel electrophoresis (1.2% TAE).
1 2 3 4 5 6 71 Perfect DNA™ Markers, 0.5–12 kbp
2 0.5 kbp PCR product
3 1.0 kbp PCR product
4 2.0 kbp PCR product
5 4.8 kbp PCR product
6 7.35 kbp PCR product
7 Perfect DNA Markers, 0.5–12 kbp
bp
12,000 –8000 –6000 –
4000 –
3000 –
2000 –
1500 –
1000 –
500 –
www.novagen.com10
NovaTaq™ PCR Kit PLUS
The NovaTaq™ PCR Kit PLUS includes 1.5 ml of 10X NovaTaq
Optimization Buffer in addition to all of the NovaTaq PCR Kit
reagents. The NovaTaq Optimization Buffer offers advantages of
greater tolerance to variable Mg2+ concentrations and a wider
temperature window for optimal primer: template annealing. Sufficient
amounts of reagents are provided for 200 standard 50 µl amplification
reactions (or 100 × 100 µl reactions).
The kit includes NovaTaq DNA Polymerase (250 U at 5 U/µl), 10X
NovaTaq Buffer with MgCl2,10X NovaTaq Buffer without MgCl2, 10X
NovaTaq Optimization Buffer, 25 mM MgCl2, 10 mM dNTP Mix and
PCR Grade Water.
ADVANTAGES
• Maximum PCR reaction versatility and optimization
• PCR optimization buffer included
NovaTaq™ PCR Master Mix is a ready-to-use 2X concentrated mixture of
NovaTaq DNA Polymerase, ultrapure deoxynucleotides, and reaction buffer
without MgCl2. The Master Mix simplifies the assembly of PCR reactions
and offers advantages of time savings, consistency, and minimal risk of con-
tamination. Simply add the NovaTaq PCR Master Mix to an equal volume
containing the required amount of MgCl2, DNA template, and primers, and
the reaction is ready for thermal cycling.The final diluted reaction contains
2.5 U of NovaTaq DNA Polymerase per 100 µl. Sufficient components are
included for 100 amplification reactions.
The kit includes NovaTaq PCR Master Mix, 25 mM MgCl2 and PCR Grade
Water.
ADVANTAGES
• Reproducibility
• Accuracy; no pipetting errors
• Minimal risk of contamination
• Convenience
NovaTaq PCR Master Mix
For ordering information, see page 25.
q u a l i t yp e r f o r m a n c e
TA Q P O LY M E R A S E S A N D PCR K I T S
NovaTaq™ Hot Start DNA Polymerase
NovaTaq™ Hot Start DNA Polymerase is a heat-activatable, chemical-
ly modified form of Taq DNA polymerase that is inactive at room tem-
perature. NovaTaq Hot Start DNA Polymerase provides improved
specificity when compared to standard Taq DNA polymerase and can
eliminate generation of non-specific amplification products such as
primer-dimers and misprimed products.The enzyme must be activat-
ed by heat treatment (7–10 min at 95˚C) before polymerization is
possible. The enzyme leaves single 3'-dA overhangs that make the
products suitable for cloning with Novagen’s AccepTor™ Vector Kits.
Each order also includes optimized 10X NovaTaq Hot Start Buffer and
a separate vial of 25 mM MgCl2.
ADVANTAGES
• Higher PCR specificity and yield
• Improved low-copy target amplification
• Automated room temperature set up
• Target amplification of up to 5 kbp
• Ideal for quantitative PCR applications
Activation Profile of NovaTaq Hot Start DNA Polymerase
A M P L I F Y
Heat-activatable modified form of recombinant Taq DNA Polymerase
Hot Start PCR products
1 Perfect DNA™ Markers, 0.05–10 kbp2 1.0 kbp DNA fragment amplified using
NovaTaq Hot Start DNA Polymerase3 1.0 kbp DNA fragment amplified using
Competitor A chemically-modified TaqDNA polymerase
1 2 3
www.novagen.com12
The Taq Antibody is a mouse monoclonal antibody that inhibits Taq
DNA polymerase activity at ambient temperatures. It provides an anti-
body-mediated hot start that enhances the specificity, sensitivity, and
convenience of PCR. Inhibition is effective during reaction assembly at
room temperature, and is completely reversed when thermal cycling
begins, with no other effect on PCR conditions.The antibody inhibits
both native and recombinant Taq DNA polymerase activities, including
NovaTaq DNA Polymerase.
One microgram (1 µl) of antibody inhibits > 95% of 5 units of Taq
DNA polymerase. For convenience, simply mix 100 µl Taq Antibody
with 500 U (100 µl) NovaTaq DNA Polymerase, incubate for 5 minutes
at room temperature, and proceed with PCR. The polymerase:anti-
body complex can be freshly prepared for each experiment or stored
at –20°C for later use.
ADVANTAGES
• Higher PCR specificity and yield
• Improved low-copy target amplification
• Room temperature set-up compatible with automation
Taq Antibody
For ordering information, see page 25.
h i g h t h r o u g h p u t
H E A T A C T I V A T A B L E
Converts unmodified Taq DNA polymerase into a hot start enzyme
SpinPrep™ DNA Purification Kits
The SpinPrep™ Kits are silica membrane-based DNA purification
systems that are easy to use and yield pure DNA quickly. For routine
clean-up of PCR products, isolation of DNA from gels, and purification
of high quality plasmid DNA, these kits offer a convenient spin column
format. Protocols do not require organic extraction or precipitation,
and the DNA is eluted in TE buffer, ready for sequencing, subcloning,
labeling, transformation, or PCR.The SpinPrep Master Kit combines
materials from the SpinPrep Plasmid Kit and SpinPrep Gel DNA Kit.
DNA P U R I F I C AT I O N
P U R I F Y
SpinPrepPlasmid Kit
Rapid, high quality plasmid minipreps
SpinPrepGel DNA Kit
Rapid, efficient extraction of DNA fromagarose gels
SpinPrepPCR Clean-up Kit
Rapid purification of PCR products fordownstream procedures
1 2 3 4 5
12,000 –
8000 –
1000 –
150 –
Gel analysis and yield of plasmids isolatedwith the SpinPrep Plasmid Kit
1 2 3 4 5 6
1 2 3 4 5 6
gel lane
18
16
14
12
10
8
6
4
2
0
µg p
lasm
id e
lute
d
bp
Lane Sample Recovery1 control mix –2 150 bp 90%3 1,000 bp 90%4 8,000 bp 50%5 12,000 bp 52%
Gel analysis and yield of DNA fragmentsisolated with the SpinPrep Gel DNA Kit
Primer removal with the SpinPrep PCRClean-up Kit
1 2 3 4bp
2000 –1500 –1000 –750 –
500 –
300 –
150 –
50 –
1 PCR Markers2 crude PCR product3 purified PCR product4 PCR Markers
1 ml culture
high copy plasmid
3 ml culture
low copy plasmid
SpinPrepMaster Kit
Materials provided for 20 plasmid
minipreps and 20 extractions from agarose gels
www.novagen.com14
For ordering information, see page 26.P u r i t ys p e e d
Product Scale DNA Yield Time Required Size Range Recommended Applications Procedure
SpinPrep™ PCR DNA in PCR Membrane binding < 10 minutes 100 bp to Ligation, labeling, sequencing,Clean-up Kit reactions capacity 6 µg > 12,000 bp PCR, transformation,
(100 µl per rxn) in vitro transcription
SpinPrep DNA in agarose Membrane binding < 20 minutes 150 bp to Ligation, labeling, sequencing,Gel DNA Kit gel slices capacity 20 µg > 12,000 bp PCR, transformation,
(150 mg per rxn) in vitro transcription
SpinPrep 1–3 ml culture Up to 20 µg < 30 minutes up to 20 kbp Sequencing, restriction digest,Plasmid Kit transformation
ADVANTAGES
• Fast and easy
• High yield
• Proven silica membrane technology
• Simple, reproducible procedure
• Alkaline lysis of E. coli (3 steps)
• Transfer supernatant to membrane. Spin.
• Add wash buffer. Spin.
• Add elution buffer. Spin.
• Dissolve gel slice (in chaotrope provided)
• Transfer to membrane. Spin.
• Add wash buffer. Spin.
• Add elution buffer. Spin.
• Layer sample on membrane. Spin.
• Add wash buffer. Spin.
• Add elution buffer. Spin.
P E L L E T PA I N T ® C O - P R E C I P I T A N T S
Non-fluorescent visible co-precipitant for automated sequencing applications
Pellet Paint NF Co-Precipitant* is a non-fluorescent dye-labeled
carrier compatible with fluorescent sequencing and other applications.
It can increase precipitation speed and efficiency, lower centrifugation
times, and help rapidly clean up unincorporated BigDye™ terminators.
Since nucleic acid pellets with Pellet Paint NF Co-Precipitant are so
visible, small pellets are unlikely to be lost and pellet dissolution is
clearly visible.
Pellet Paint NF Co-Precipitant is fully compatible with the ABI PRISM®
BigDye Terminator Cycle Sequencing Ready Reaction. To avoid extra
sample handling, Pellet Paint NF Co-Precipitant can be added directly
to the reaction mix, template DNA, crude PCR samples, or dilution
buffer prior to the cycle sequencing reaction. Pellet Paint NF Co-
Precipitant has no detectable effect on the sequencing reaction or
sequencing accuracy.
Pellet Paint NF Co-Precipitant
Rapid, quantitative precipitation of DNA and RNA, including PCR clean-up
Pellet Paint® Co-Precipitant* is a visible dye-labeled carrier
formulated specifically for use in alcohol precipitation of nucleic acids.
The five-minute precipitation protocol requires no low temperature
incubations or prolonged centrifugation. Both RNA and DNA are
efficiently precipitated from solutions as dilute as 2 ng/ml, and the
pellet is easily located by its vivid pink color.The pellet can be easily
followed during washing steps, preventing losses during handling. Pellet
Paint is compatible with most molecular biology procedures and is free
of contaminating nucleic acids and nucleolytic enzymes. Pellet Paint is
compatible with Cy5-based automated sequencers. Pellet Paint NF
(below) is recommended for use with PE Applied Biosystems
automated sequencers.
Pellet Paint Co-Precipitant
P U R I F Y
ADVANTAGES
• Visibility maximizes sample recovery
• Suitable for DNA and RNA precipitation
• Compatible with ethanol and isopropanol precipitation
• Compatible with sodium acetate, sodium chloride and
ammonium acetate salts
• Room temperature incubation and short centrifugation times
• Preparations free of contaminating nucleic acids and protein
• Improves yield, reproducibility, and sensitivity in many
applications (e.g. PCR)
• Works even with minute quantities of nucleic acid
(< 2 ng/ml)
• Compatible with numerous downstream applications
• Pellet Paint NF Co-Precipitant compatible with BigDye
terminators and other fluorescent applications
*patent pending
BigDye and ABI PRISM are trademarks of the PE Corporation
www.novagen.com16
Add 2 µl Pellet Paint or Pellet Paint NF
Co-Precipitant + 0.1 volume 3 M Na Acetate
to sample and mix briefly
Add 2 volumes ethanol (or 1 volume isopropanol)
and briefly vortex
Incubate at room temperature for 2 minutes
Spin sample for 5 minutes
Discard supernatant, wash and resuspend pellet
PELLET PAINT® PROCEDURE
1
2
3
4
5
For ordering information, see page 26.
U S E I T so you don’t lose it
C L O N I N G K I T S
The Perfectly Blunt® Cloning Kits are designed for simplified cloning of
DNA generated by PCR using any type of DNA polymerase. This
approach enables the use of high fidelity proofreading enzymes for
amplification, thus decreasing the probability of generating mutations in
the target sequence. With the Perfectly Blunt cloning protocol, you can
go from PCR product to plating transformants in less than 1 hour with
minimal hands-on time.
The finished PCR reaction product is converted to a blunt, phosphorylat-
ed form in a 15-minute reaction using premixed reagents. Following a 5-
minute heat inactivation step, the treated insert is combined with the
ready-to-use vector and ligated in an optimized reaction using premixed
reagents. An exclusive 8-minute transformation procedure using highly
efficient NovaBlue Singles™ Competent Cells generates recombinant
colonies that are easily visualized by blue/white screening.
Six different plasmid vectors are available in Perfectly Blunt Cloning Kits.
Vector choices include those designed for general cloning, sequencing,
optimal transcription/translation, and optimal protein expression in E. coli.
Please refer to the table on the facing page for features and applications of
the vectors.
ADVANTAGES
• Blunt methodology is 3- to 24-fold more efficient than T-vectorcloning methods (1)
• No restriction enzymes or special primers
• Direct ligation of PCR product with vector
• Compatible with any DNA polymerase
• Blue/white screening with all vectors
• Simple protocol takes as little as 45 minutes from PCR product toplating transformants
Perfectly Blunt® Cloning Kits
AccepTor™ Vector Kits*
The AccepTor™ Vector Kits are designed for simplified cloning of
PCR products generated using non-proofreading thermostable DNA
polymerases, such as native and recombinant Taq polymerase, that
leave single 3'-dA overhangs on the reaction products.The linearized
AccepTor Vector contains single 3'-dU DNA ends that are
compatible with direct ligation of these products without the need for
intermediate reactions.The dU residues are converted to dT residues
in vivo following transformation.
*AccepTor Vector Kits are covered under U.S. Patent No. 5,856,144 issued to Novagen, Inc.
C L O N E
Less than 1 hour from PCR product to plating
Simplified cloning of DNA generated by PCR using any type of DNA polymerase
Rapid, direct cloning of DNA amplified with non-proofreading DNA polymerases
ADVANTAGES
• No restriction enzymes or special primers
• Compatible with polymerases that leave single 3'-dA overhangs
• Simple protocol takes less than 40 minutes from PCR product to plating transformants
• Blue/white screening with pSTBlue-1 and pETBlue™-1 vectors
1. Novy, R.E., Yaeger, K.W. and Kolb, K.M. (1996) inNovations 6, 7–11
www.novagen.com18lacZ lacZ
dA
dA
vectorvectorPCR product
dU
dU
P
PlacZ lacZ
vector vectorPCR product
F A S T
s i m p l eFor ordering information, see page 26
Vectors Applications Vector Advantages
pSTBlue-1 Archiving, Subcloning, Dual opposed SP6/T7 promotersSequencing, Amp or Kan selectionIn vitro transcription Dual EcoR I sites flank insert
pETBlue™-1 Protein expression: No fusion tagsT7lac-driven, tightly Insert provides ATG start codoncontrolled, high levelexpression in E. coli
Vectors Applications Vector Advantages
pSTBlue-1 Archiving, Subcloning, Dual opposed SP6/T7 promotersSequencing, Amp or Kan selectionIn vitro transcription Dual EcoR I sites flank insert
pT7Blue-3 T7 promoterAmp or Kan selectionDual EcoR I sites flank insert
pT7Blue T7 promoterNde I/BamHI sites flank insert
pT7Blue-2 Protein expression: T7-driven in vitro protein synthesisIn vitro transcription/translation, N-terminal S•Tag™ sequenceSequencing Optimal Kozak translation initiation
Xenopus globin 5' UTR
pETBlue-1 Protein expression: T7lac-driven No fusion tagstightly controlled, high level Insert provides ATG start codonexpression in E. coli
pETBlue-2 Optional C-terminal HSV•Tag®, His•Tag®
Vector provides ATG start codon
Perfectly Blunt® Vectors
AccepTor™ Vectors
C O M P E T E N T C E L L S
NovaBlue is an E. coli K-12 strain ideally suited as an initial cloning host
due to its high transformation efficiency, blue/white screening
capability, and recA endA mutations, which result in high yields of excel-
lent quality plasmid DNA. The NovaBlue Singles™ Competent Cells
format is designed for ultimate convenience and reliability in plasmid
transformation. Cells are provided in single use aliquots that eliminate
the need to subaliquot, freeze/thaw or waste partially used vials, thus
saving time and increasing performance.To use, simply thaw, add DNA,
incubate 5 minutes on ice, heat shock for 30 seconds, place on ice for
2 minutes and plate directly (when selecting for ampicillin resistance)
or after incubation at 37°C for 30 minutes (when selecting for
kanamycin resistance).
NovaBlue Singles Competent Cells are provided as frozen 50 µl
aliquots, each intended for one transformation, and also include Test
Plasmid and SOC Medium for the specified number of transformation
reactions.
ADVANTAGES
• Convenient, one tube per transformation. No
measuring, subdividing, shortage, waste or ß-ME
• Simple, reliable
• Cost-effective without leftovers
• Consistently high transformation efficiency
• Transformation in as little as 8 minutes
• Easy-open flip caps
NovaBlue Singles™ Competent Cells
The HT96™ NovaBlue Competent Cells are designed for high
throughput cloning. The Cells are pre-dispensed in a robust 96-well
polypropylene plate, which is compatible with a variety of thermal
cyclers as well as water baths for performing the transformation reac-
tion.The wells are individually sealed and have raised rims to prevent
cross-contamination. The seal may be easily pierced with standard
pipet tips or peeled off for access. Strips of caps are also
provided for reliable sealing during manipulation and storage. Groups
of 24 wells can by simply detached from the whole plate for process-
ing smaller numbers of samples.
HT96 NovaBlue Competent Cells are provided as 20 µl aliquots in a
96-well plate. Each package also includes cap strips, SOC Medium,Test
Plasmid and a Reagent Reservoir.
ADVANTAGES
• Seal may be pierced or peeled off
• Cap strips provided for resealing
• Plate may be separated into smaller groups of wells
• Compatible with many thermal cyclers
HT96™ NovaBlue Competent Cells
T R A N S F O R M
Premeasured, ready-to-use aliquots for speedy, hassle-free transformation
High efficiency competent cells pre-dispensed in a 96-well format
www.novagen.com20
For ordering information, see page 27.
3 S t e p sT h a t ’ s i t !
1 5 M i n u t e s
High throughput cloning usingHT96 Competent Cells
HT96™ Isothermal BlockEfficient thermal transfer for uniform incubations
The HT96™ Isothermal block is an anodized aluminum, solvent-resistant
block specially designed to hold HT96 plates. It provides efficient thermal
transfer during the low temperature incubation and heat-shock steps
used in transformation protocols. Simply pre-incubate the anodized alu-
minum block at the desired temperature and place the HT96 Competent
Cell plate in the block. The overall block dimensions are
12 cm x 7.6 cm x 2.5 cm and it is compatible with most 96 well PCR
plates. The HT96 plate is shown in the HT96 Isothermal Block in the
photo below.
Blue/white screening withNovaBlue Competent Cells
DNA L A D D E R S A N D M A R K E R S
Perfect DNA™ Ladders and Markers are available in several conven-
ient size ranges for gel analysis.The DNA species are supplied in equal
masses, except for a 2–3X reference band in some marker sets. Each
marker or ladder is supplied in 1X DNA Gel Loading Buffer. A
separate vial of 6X DNA Gel Loading Buffer is included.
ADVANTAGES
• Ready-to-use mixtures
• Evenly spaced, easy-to-remember sizes
• Includes gel loading buffer
• Consistent lot-to-lot, quality-assured product ensures
reproducible results
• Available in various size ranges
Perfect DNA™ Ladders and Markers
A N A L Y Z E
www.novagen.com22
bp– 3000
– 2000
– 1000
– 900– 800– 700
– 600
– 500– 450– 400– 350– 300– 250
– 200
– 150
– 100
– 50
bp
– 1000– 900– 800– 700– 600
– 500
– 400
– 300
– 200
– 100
50 bp Ladder2.0% TAE agarose gel
100 bp Ladder2.0% TAE agarose gel
1 kbp Ladder0.8% TAE agarose gel
bp
– 10,000– 8000– 6000– 5000– 4000– 3000– 2500 – 2000
– 1500
– 1000
– 500
bp
– 12,000 – 10,000– 8000
– 6000
– 4000*
– 3000
– 2000
– 1000
– 1500
– 500
0.5–12 kbp0.8% TAE agarose gel
0.1–12 kbp1.5% TAE agarose gel
bp– 12,000– 10,000– 8000– 6000
– 4000*
– 3000
– 2000
– 1500
– 1000
– 500* – 400– 300– 200– 100
bp– 10,000– 8000– 6000– 4000– 3000
– 2000
– 1500– 1400
– 1000
– 750
– 500
– 400
– 300
– 200
– 100
– 50
0.05–10 kbp2.0% TAE agarose gel
Perfect DNA Ladders
Perfect DNA Markers
p e r f e c t l y a c c u r a t ew i d e r a n g e
For ordering information, see page 27.
PCR Markers
The PCR Markers are a mixture of eight defined DNA fragments for
characterizing small DNA products. The markers are supplied in gel
loading buffer containing two tracking dyes that do not interfere with
UV illumination of ethidium bromide-stained bands. A separate vial
of 6X Loading Buffer is included. The recommended amount for
loading produces bands of even intensity that are bright, sharp, and
easy to photograph.
ADVANTAGES
• Ready-to-use mixture
• Accurate sizing of PCR poducts
• Includes gel loading buffer
• Consistent lot-to-lot, quality-assured product ensures
reproducible results
50–2000 bp1.5% TAE agarose gel
bp
– 2000
– 1500
– 1000
– 750
– 500
– 300
– 150
– 50
O R D E R I N G I N F O R M AT I O N
Prices subject to change
24
PRODUCT SIZE CAT NUMBER PRICE
KOD HIFI DNA POLYMERASE * 250 U 71085-3 £148COMPONENTS:
• 250 U KOD HiFi DNA Polymerase (2.5 U/µl )• 1 ml 10X KOD HiFi DNA Polymerase PCR Buffer #1• 1 ml 10X KOD HiFi DNA Polymerase PCR Buffer #2• 1 ml 2 mM dNTP Mix• 1 ml 25 mM MgCl2
KOD HOT START DNA POLYMERASE * 200 U 71086-3 £1285 × 200 U 71086-4 £480
COMPONENTS:• 200 U or 5 × 200 U KOD Hot Start DNA Polymerase (2.5 U/µl)• 1.2 ml or 5 × 1.2 ml 10X KOD Hot Start PCR Buffer• 1 ml 25 mM MgSO4
• 1 ml or 5 × 1 ml 2mM dNTP Mix
KOD XL DNA POLYMERASE * 250 U 71087-3 £1655 × 250 U 71087-4 £640
COMPONENTS:• 250 U or 5 × 250 U KOD HiFi DNA Polymerase (2.5 U/µl )• 1 ml or 5 × 1 ml 10X KOD XL DNA Polymerase PCR Buffer • 1 ml or 5 × 1 ml 2 mM dNTP Mix
GENOMIC DNA
Bovine blood 69231-3 £80Cat blood 69235-3 £80Chicken blood 69233-3 £80Dog blood 69234-3 £80Human (female only) blood 70605-3 £80Human (male only) blood 70572-3 £80Human (male & female) blood 69237-3 £80Mouse blood 69239-3 £80Pig blood 69230-3 £80Rat (Sprague Dawley) blood 69238-3 £80S. cerevisiae (wild type S288C) mixed culture 69240-3 £80
All DNAs are supplied at 100–300 µg/ml in TE buffer, 100 µg per vial.
FIRST STRAND CDNA SYNTHESIS KIT 40 rxn 69001-3 £159COMPONENTS: • 20 µg Oligo(dT) Primer
• 4000 U MMLV Reverse Transcriptase • 10 µg Random Hexamer Primers• 0.2 ml 5X First Strand Buffer • 1.5 ml Nuclease-free Water• 0.1 ml 100 mM DTT • Positive Control RNA • 50 µl 10 mM dNTP Mix • Positive Control Primers
ONE STEP RT-PCR KIT * 50 rxn 71089-3 £176COMPONENTS:
• 250 U rTth DNA Polymerase (2.5 U/µl) • 2 × 1.1 ml RNase Free Water• 1 ml 5X Reaction Buffer • 0.05 ml 10 mM G3PDH Control Primer F • 0.5 ml 25 mM Mn(OAc)2 • 0.05 ml 10 mM G3PDH Control Primer R• 0.3 ml 2.5 mM dNTP Mix • 0.05 ml Positive control Human G3PDH • 0.1 ml RNase Inhibitor (10 U/µl) RNA (5 x 108 copies/ml)
25
Prices subject to change* These products are not available through Novagen in Japan
Purchase of NovaTaq DNA Polymerase enzymes and kits, KOD DNA Polymerase enzymes and kits, and the One Step RT-PCR Kit are accompanied by a limited license to use these products in the Polymerase Chain Reaction (PCR) process for research use in conjunction with a thermal cyclerwhose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Applied Biosystems or as purchased,i.e., an authorized thermal cycler.
KOD DNA Polymerases, the One Step RT-PCR Kit and Taq Antibody are manufactured by Toyobo and distributed by Novagen.
KOD XL DNA Polymerase is licensed under US Patent Number 5,436,149 owned by Takara Shuzo Co., Ltd.
PRODUCT SIZE CAT NUMBER PRICE
NOVATAQ™ DNA POLYMERASE 100 U 71003-3 £38500 U 71003-4 £1652,500 U 71003-5 £759
ADDITIONAL COMPONENTS:• 1 or 2 or 7 × 1.5 ml 10X NovaTaq Buffer with MgCl2• 1 or 2 or 7 × 1.5 ml 10X NovaTaq Buffer without MgCl2• 1 or 2 or 7 × 1.5 ml 25 mM MgCl2
10 MM DNTP MIX 0.2 ml 71004-3 £33
NOVATAQ PCR KIT 250 U 71005-3 £119COMPONENTS:
• 250 U NovaTaq DNA Polymerase (5 U/µl) • 1.5 ml 25 mM MgCl2• 1.5 ml 10X NovaTaq Buffer with MgCl2 • 0.2 ml 10 mM dNTP Mix• 1.5 ml 10X NovaTaq Buffer without MgCl2 • 5 × 2ml PCR Grade Water
NOVATAQ PCR KIT PLUS 250 U 71006-3 £127COMPONENTS:
• 250 U NovaTaq DNA Polymerase (5 U/µl) • 1.5 ml 25 mM MgCl2• 1.5 ml 10X NovaTaq Buffer with MgCl2 • 0.2 ml 10 mM dNTP Mix• 1.5 ml 10X NovaTaq Buffer without MgCl2 • 5 × 2ml PCR Grade Water• 1.5 ml 10X NovaTaq Optimization Buffer
NOVATAQ PCR MASTER MIX 250 U 71007-3 £119COMPONENTS:
• 4 × 1.25 ml 2X NovaTaq PCR Master Mix• 1.5 ml 25 mM MgCl2• 3 × 2 ml PCR Grade Water
NOVATAQ HOT START DNA POLYMERASE 250 U 71091-3 £855 x 250 U 71091-4 £370
COMPONENTS:• 250 U or 5 x 250 U NovaTaq Hot Start DNA Polymerase (5 U/µl)• 1 or 5 × 1.5 ml 10X NovaTaq Hot Start Buffer• 1 or 5 × 1.5 ml 25 mM MgCl2
TAQ ANTIBODY * 100 µl 71088-3 £128COMPONENTS:
• 100 µl Taq Antibody (concentration 1µg/µl)• 1 ml 10X PCR Buffer
26
O R D E R I N G I N F O R M AT I O N
PRODUCT SIZE CAT NUMBER PRICE
SPINPREP™ PCR CLEAN-UP KIT 100 rxn 70976-3 €106INTRO. SPINPREP PCR CLEAN-UP KIT 20 rxn 70975-3 €26
SPINPREP GEL DNA KIT 100 rxn 70852-3 €106INTRO. SPINPREP GEL DNA KIT 20 rxn 70958-3 €26
SPINPREP PLASMID KIT 100 rxn 70851-3 €106INTRO. SPINPREP PLASMID KIT 20 rxn 70957-3 €26
SPINPREP MASTER KIT 40 rxn 71073-3 €58
PELLET PAINT® CO-PRECIPITANT 125 rxn 69049-3 €60PELLET PAINT NF CO-PRECIPITANT 125 rxn 70748-3 €60
COMPONENTS:• 250 µl Pellet Paint or Pellet Paint NF Co-Precipitant• 1 ml 3 M Sodium Acetate, pH 5.2
PSTBLUE-1 PERFECTLY BLUNT® CLONING KITS 10 rxn 70184-3 €10520 rxn 70191-3 €196
40 rxn 70191-4 €327
PT7BLUE-3 PERFECTLY BLUNT CLONING KITS 10 rxn 70075-3 €10520 rxn 70182-3 €19640 rxn 70182-4 €327
PT7BLUE-2 PERFECTLY BLUNT CLONING KITS 10 rxn 70185-3 €10520 rxn 70190-3 €19640 rxn 70190-4 €327
PT7BLUE PERFECTLY BLUNT CLONING KITS 10 rxn 70183-3 €10520 rxn 70189-3 €19640 rxn 70189-4 €327
PETBLUE™-1 PERFECTLY BLUNT CLONING KITS 10 rxn 70633-3 €11620 rxn 70634-3 €21940 rxn 70634-4 €360
PETBLUE-2 PERFECTLY BLUNT CLONING KITS 10 rxn 70635-3 €11620 rxn 70636-3 €21940 rxn 70636-4 €360
PSTBLUE-1 ACCEPTOR™ VECTOR KITS 10 rxn 70594-3 €10520 rxn 70595-3 €19640 rxn 70595-4 €327
PETBLUE-1 ACCEPTOR VECTOR KITS 10 rxn 70597-3 €11620 rxn 70598-3 €21940 rxn 70598-4 €360
27For a complete listing of kit components, please refer to Novagen’s website: www.novagen.com
PRODUCT SIZE CAT NUMBER PRICE
NOVABLUE SINGLES™COMPETENT CELLS 11 rxn 70181-3 €8622 rxn 70181-4 €165
COMPONENTS:• 11 × 50 µl or 22 × 50 µl Competent Cells• 10 µl Test Plasmid• 2 × 2 ml or 4 × 2 ml SOC Medium
HT96™ NOVABLUE COMPETENT CELLS 1 plate 71011-3 €4944 plates 71011-4 €180420 plates 71011-5 €6872
COMPONENTS:• 96 × 20 µl or 4 × (96 × 20µl) Competent Cells
or 20 × (96 × 20µl)• 10 µl or 2 × 10 µl or 10 × 10 µl Test Plasmid• 14 ml or 4 × 14 ml or 20 × 14 ml SOC Medium• pkg/l2 or 4 × pkg/l2 or 20 × pkg/l2 8 Cap Strip• 1 or 4 or 20 Reagent Reservoir
HT96 ISOTHERMAL BLOCK 1 ea 71031-3 €84
PERFECT DNA™ 50 BP LADDER 100 lanes 70538-3 €99PERFECT DNA 100 BP LADDER 100 lanes 70539-3 €99PERFECT DNA 1 KB LADDER 100 lanes 70537-3 €99
PERFECT DNA MARKERS, 0.5–12 KBP 100 lanes 69002-3 €99PERFECT DNA MARKERS, 0.1–12 KBP 100 lanes 70087-3 €99PERFECT DNA MARKERS, 0.05–10 KBP 100 lanes 70540-3 €99
PCR MARKERS, 50–2000 BP 50 lanes 69278-3 €75
All Markers include 6X Loading Buffer
Prices subject to change
CN Biosciences (UK) Ltd. VWR International LtdBoulevard Industrial Park Hunter BoulevardPadge Road, Beeston Magna ParkNottingham NG9 2JR Lutterworth LE17 4XN
Tel. 0115 9430840 Freephone 0800 223344Fax 0115 9430951 Fax 01455 558586e-mail [email protected] Sales e-mail [email protected] www.cnuk.co.uk Web www.vwr.com
Calbiochem-Novabiochem GmbH VWR International GmbHOber der Röth 4, 65824 Schwalbach/Ts. D-64271 Darmstadt
Ordering:
Freecall 0800 69 31 000 Tel. 0180 570 2000Freefax 0800 62 36 100 Fax 0180 570 2222e-mail [email protected] e-mail [email protected] www.cnbi.de
Technical Service:
Freecall 0800 6100 34 96e-mail [email protected]
Novagen is a brand of An Affiliate of Merck KGaA, Darmstadt, Germany
Novagen, Inc. VWR International 601 Science Drive Tel. 800 932 5000Madison,WI 53711 Fax 800 668 6348
Web www.vwr.comFax 608 238 1388Tel. 608 238 6110Tel. 800 526 7319e-mail [email protected] www.novagen.com
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