rhAmp Genotyping Master Mix and Reporter Mixes
CAT# PRODUCT SIZE STORAGE Һڰ ֹ
1076014[IDT] rhAmp Genotyping Master Mix1 X 0.5 mL-α
1076015[IDT] rhAmp Genotyping Master Mix1 x 5 mL-α
1076016[IDT] rhAmp Genotyping Master Mix2 x 5 mL-α
1076017[IDT] rhAmp Genotyping Master Mix5 x 5 mL-α
1076018[IDT] rhAmp Genotyping Master Mix1 x 50 mL-α
1076020[IDT] rhAmp Reporter Mix w/Reference 1 x 25 uL-α
1076021[IDT] rhAmp Reporter Mix w/Reference 1 x 250 uL-α
1076022[IDT] rhAmp Reporter Mix w/Reference 1 x 500 uL-α
1076023[IDT] rhAmp Reporter Mix w/Reference1250 uL-α
1076024[IDT] rhAmp Reporter Mix w/Reference1 x 1250 uL-α
1076025[IDT] rhAmp Reporter Mix 1 x 25 uL-α
1076026[IDT] rhAmp Reporter Mix 1 x 250 uL-α
1076027[IDT] rhAmp Reporter Mix 1 x 500 uL-α
1076028[IDT] rhAmp Reporter Mix1250 uL-α
1076029[IDT] rhAmp Reporter Mix 1 x 2500 uL-α

Amplification and reporter mixes optimized for rhAmp SNP Genotyping

rhAmp Genotyping Master Mix and rhAmp Reporter Mixes are specially formulated for use with rhAmp SNP Assays to deliver superior specificity and signal-to-noise ratios for high confidence genotype calls. rhAmp Genotyping Master Mix is a unique, two-enzyme formulation optimized for highly specific amplification and accurate allele discrimination. rhAmp Reporter Mixes contain a universal 5 nuclease reporter system to distinguish the reference allele from the alternative allele for any rhAmp SNP Assay.

  • Easily report high accuracy calls with novel RNase H2 assay chemistry 
  • Generate high confidence calls using Hawkeye Taq DNA Polymerase with enhanced allelic discrimination
  • Eliminate expensive custom probe synthesis with cost-effective universal reporter probes
  • Use with any common commercial real-time PCR instrument by selecting mix with or without reference dye


rhAmp Genotyping Master Mix

A 2X master mix solution, containing the required enzymes and additional components necessary for activation and amplification of rhAmp SNP Assays.
Product Name
(Cat #)
Size
Һڰ
rhAmp Genotyping Master Mix
1076014
1 X 0.5 mL
       86,000
1076015
1 x 5 mL
     478,000
1076016
2 x 5 mL
     849,000
1076017
5 x 5 mL
   1,713,000
1076018
1 x 50 mL
   3,307,000


rhAmp Reporter Mix

A 40X universal, reporter probe mix for use with rhAmp Genotyping Master Mix and rhAmp SNP Assays. Does not contain reference dye.
Product Name
(Cat #)
Size
Һڰ
rhAmp Reporter Mix
1076025
 1 x 25 µL 
       29,000
1076026
 1 x 250 µL 
       77,000
1076027
 1 x 500 µL 
     117,000
1076028
  1250 µL 
     183,000
1076029
 1 x 2500 µL 
     338,000


rhAmp Reporter Mix w/Reference

A 40X universal, reporter probe mix with reference dye required by certain instruments. For use with rhAmp Genotyping Master Mix and rhAmp SNP Assays.
Product Name
(Cat #)
Size
Һڰ
rhAmp Reporter Mix w/Reference
1076020
 1 x 25 µL 
       33,000
1076021
 1 x 250 µL 
       86,000
1076022
 1 x 500 µL 
     129,000
1076023
  1250 µL 
     204,000
1076024
 1 x 2500 µL 
     375,000


rhAmp SNP Genotyping technology incorporates a novel method using blocked PCR primers that contain a single RNA base. The rhAmp primers are activated when the blocking motif is removed by the RNase H2 in the rhAmp Genotyping Master Mix. Cleavage occurs at the 5 end of the RNA base (Figure 1) only when it is perfectly matched to its DNA complement. Primer cleavage occurs in the background during each anneal/extend cycle to generate a 3 hydroxyl for polymerase extension, which means the PCR protocol does not require any additional steps. The mechanism creates extremely tight control of amplification that greatly reduces primer-dimers and off-target amplification of closely related sequences.


RNA Base cleavage (Structures)

Figure 1. RNase H2 cleavage produces a 3-hydroxyl group for extension by Hawkeye Taq Polymerase. RNase H2 cleaves the phosphate backbone between the DNA base and RNA base during normal PCR cycling conditions. This produces a 3-hydroxyl group on the DNA base that is accessible to DNA polymerase extension.

 

rhAmp Genotyping Master Mix

rhAmp Genotyping Master Mix contains hot start versions of RNase H2 from Pyrococcus abyssi (P.a.), and a novel Hawkeye Taq DNA Polymerase with enhanced sensitivity for allelic mismatches. This dual enzyme formulation, combined with rhAmp SNP Assays, provides highly sensitive, allele-specific PCR for the detection of single-nucleotide polymorphisms.

Pyrococcus abyssi is an extreme thermophile. Therefore, P.a. RNase H2 enzyme has optimal activity between 70C and 75C and is functional in rhPCR between 50C and 75C. P.a. RNase H2 has very low activity at room temperature (~1000X less active). These properties allow the enzyme to deliver excellent performance in the optimized rhAmp Genotyping PCR buffer conditions.

The Hawkeye Taq DNA Polymerase is a novel, proprietary polymerase that is optimized for allelic discrimination. The Hawkeye Taq polymerase and RNase H2 combination delivers clear discrimination and high signal-to-noise ratios, due in part to reagents not being unnecessarily consumed by primer-dimers and spurious amplification products.

rhAmp Reporter Mix

rhAmp Reporter Mix is a cost-effective, universal, 5 nuclease assay reporter mix that contains 2 universal probes to distinguish the reference allele from the alternate allele and a universal forward primer. The FAM universal probe is assigned to the reference allele, and the Yakima Yellow universal probe is assigned to the alternate allele. The Yakima Yellow reporter dye can be read on qPCR instruments in the VIC channel without need for calibration.

The universal primer and probe designs have been optimized to deliver consistent, robust signal-to-noise for improved automated allele calling. And, unlike traditional 5 nuclease SNP assays, the rhAmp SNP universal reporter system eliminates the expense of synthesizing two SNP-specific probes for every assay. This system enables rhAmp SNP Assays to provide higher quality genotypes at a more affordable price.

  • rhAmp Reporter Mix sizes are configured to complement rhAmp Genotyping Master Mix product sizes for easy planning and ordering (Table 1)
  • Available with or without reference dye for maximum flexibility on all qPCR platforms (Table 2)

Table 1. rhAmp Reporter Mix, with or without reference dye, matched to correct size rhAmp Genotyping Master Mix.

rhAmp Reporter Mix*
Or
rhAmp Reporter Mix w/Reference dye

rhAmp Genotyping Master Mix matching size

Reactions

25 L

0.5 mL (1 X 0.5 mL)

100

250 L

5.0 mL (1 X 5 mL)

1000

500 L

10 mL (2 X 5 mL)

2000

1250 L

25 mL (5 X 5 mL)

5000

2500 L

50 mL (1 X 50 mL)

10,000

* For use with real-time qPCR instruments that do not require reference dye.
For use with real-time qPCR instruments that require reference dye.
Number of reactions based on 10 µL reaction volume.

Table 2. Reference dye requirements for various PCR systems. For instruments not listed, please check with the manufacturer.

 

Reference dye required?

PCR system

Yes

No

7900HT Fast and 7300 Real-Time PCR System (Thermo Fisher Scientific)

X

 

StepOne and StepOnePlus Real-Time PCR System (Thermo Fisher Scientific)

X

 

Mx3005P and Mx4000P qPCR System (Agilent)

X

 

7500 Real-Time PCR System (Thermo Fisher Scientific)

X

 

Viia7 Real-Time PCR System (Thermo Fisher Scientific)

X

 

QuantStudio Flex Systems (Thermo Fisher Scientific)

X

 

CFX, iQ, and Opticon Real-Time PCR Detection Systems (Bio-Rad)

 

X

LightCycler Real-Time PCR Systems (Roche)

 

X

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