KAPA HRM Fast PCR Kit (91 min) MeltDoctor™ HRM Master Mix (132 min)

Unless otherwise stated in references, presented data on file For Research Use Only. Not for use in diagnostic procedures.

Accurate detection of DNA sequence variations.

KAPA HRM FAST

High Resolution Melt (HRM) analysis is a technique for fast, high-throughput post-PCR analysis of genetic mutations or variance in nucleic acid sequences. The KAPA HRM FAST Master Mix contains a novel DNA polymerase and buffer system, and EvaGreen® saturating dye optimized for maximum discrimination between sequence variants.

Kits offer:

• SNP genotyping• mutation discovery (gene scanning)• screening for heterozygosity• DNA fingerprinting• DNA methylation analysis

Accurately Detect Type IV SNPs• 95% detection levels, even when competitors fail*

• Reduced reaction times

Allelic Discrimination Plot

*A) The Type IV SNP rs641805 is located on an intron of the dihydropyrimidine dehydrogenase (DPD) gene. Reactions also included 2.5 mM MgCl2, 0.2 μM of each primer and 10 ng of human genomic DNA. PCR and HRM was performed using a fast, 2-step cycling protocol (40 cycles) with 5 sec denaturation (95°C) and 20 sec annealing/extension (60°C).

*B) The Type IV SNP rs641805 is located on an intron of the dihydropyrimidine dehydrogenase (DPD) gene. Reactions also included 2.5 mM MgCl2, 0.2 μM of each primer and 10 ng of human genomic DNA. PCR and HRM was performed using a fast, 2-step cycling protocol (40 cycles) with 5 sec denaturation (95°C) and 20 sec annealing/extension (60°C). The ABI MeltDoctor HRM Master Mix was used according to the manufacturer’s instructions, with the same template DNA samples.

KAPA HRM FAST PCR Kit (91 min)

Temperature (ºC)

Nor

mal

ized

min

us A

/A

74.5 75 75.5 76 76.5 77 77.5 78 78.5 79 79.5

-16

-14

-12

-10

-8

-6

-4

-2

0

2

4MeltDoctor™ HRM Master Mix (132 min)

Temperature (ºC)

Nor

mal

ized

min

us A

/A

73 73.5 74 74.5 75 75.5 76 76.5 77 77.5 78

-16

-14

-12

-10

-8

-6

-4

-2

0

2

4A B

DNA extracts were prepared from buccal swabs obtained from 36 individuals using KAPA Express Extract DNA Extraction Kits (according to recommended protocols). DNA extracts were used directly in reactions also containing 2.5 mM MgCl2 and 0.2 μM of each primer. PCR and HRM was performed using a fast, 2-step cycling protocol (45 cycles) with 5 sec denaturation (95°C) and 30 sec annealing/extension (60°C).

KAPA HRM Fast PCR Kit (90 min) Type-it® HRM™ Kit (115 min)SsoFast™ EvaGreen®

Supermix (92 min)

KapaBiosystemsKapaBiosystems@KapaBiosystems

Unless otherwise stated in references, presented data on file© 2016 Kapa Biosystems. All trademarks are the property of their respective owners. SS203001 A191 10/16For Research Use Only. Not for use in diagnostic procedures.

Headquarters, United StatesWilmington, MassachusettsTel: 781.497.2933Fax: 781.497.2934

International OfficeCape Town, South AfricaTel: +27.21.448.8200 Fax: +27.21.448.6503

Kapa Technical Supportkapabiosystems.com/support

Kapa Salessales@kapabiosystems.com

Go from Cell to SNP Quickly• Enables extraction, amplification and HRM in less than 2 hours when used in combination with the KAPA Express Extract Kit

Ordering InformationRoche Cat. No. Kapa Code Description Kit Size

07959290001 KK4201 KAPA HRM Fast PCR Kit 1 mL

07959303001 KK4202 KAPA HRM Fast PCR Kit 5 mL

07959311001 KK4203 KAPA HRM Fast PCR Kit 10 mL

Temperature (ºC)

Nor

mal

ized

min

us A

/A

70.5 71 71.5 72 72.5 73 73.5 74 74.5 75 75.5 76 76.5 77 77.5 78 78.5 79

-8

-6

-4

-20

2

4

68

10

12

1416

18

20

222426

Maximize Differences in Sequence Variants

The Type I SNP rs12913832 is associated with expression of the OCA2 gene, and part of the eye color haplotype. Reactions also included 2.5 mM MgCl2, 0.2 µM of each primer and 10 ng of human genomic DNA. PCR and HRM was performed using a fast, 2-step cycling protocol (40 cycles) with 5 sec denaturation (95°C) and 20 sec annealing/extension (60°C). Competitor kits were used according to manufacturers’ instructions, with the same template DNA samples.

• Engineered to maximize differences in melting behavior

• Improved sensitivity

KAPA HRM FAST PCR Kit (90 min)

Temperature (ºC)

Nor

mal

ized

min

us A

/A

72 73 74 75 76 77 78 79 80 81 82

-6

-4

-2

0

2

4

6

8

10

12

14

16

18

20

22

24

Type-it® HRM™ Kit (115 min)

Temperature (ºC)N

orm

aliz

ed m

inus

A/A

72 73 74 75 76 77 78 79 80 81 82

-6

-4

-2

0

2

4

6

8

10

12

14

16

18

20

22

24SsoFast™ EvaGreen® Supermix (92 min)

Temperature (ºC)

Nor

mal

ized

min

us A

/A

70 71 72 73 74 75 76 77 78 79 80 81

-6

-4

-2

0

2

4

6

8

10

12

14

16

18

20

22

24