KAPA2G Fast HotStart PCR Mix is Great for Running Large Numbers of Genotyping PCRs

November 17, 2015

Stanford
Pathology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Kapa Biosystems

Product Name:

KAPA2G Fast PCR HotStart ReadyMix with dye, 500 x 25 µL reactions

Catalog Number:

KK5609

Our laboratory utilizes a wide variety of mouse models, necessitating a large amount of genotyping. The KAPA2G PCR mix is great for this use because of the great convenience (just need to add primers and DNA) and the faster PCR program (allowing genotyping to be completed more quickly)

Experimental Design and Results Summary

Application

Genotyping PCRs

Starting Material

The KAPA2G PCR mix, mouse tail DNA, and genotyping primers

Protocol Overview

The KAPA mix (2X) is added to strip tubes with the proper mouse tail DNA and genotyping primers in them, then run using the program specified in the KAPA2G data sheet.

Tips

The PCR mix can be used at 3.75 uL (for a total PCR reaction volume of 7 uL) and still work fine.

Results Summary

The KAPA PCR mix results in easily interpretable genotyping results (even with the faster program and scaled down volume).

Additional Notes

One downside is that some of my genotyping primers do not seem to work with the KAPA mix unless the program is extended to the length of a normal PCR program (longer denaturation, annealing, and melting times). But with the extended program, the PCR seems to work fine.

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Summary

The Good

Fast PCR program, very convenient to use (mix is already ready), and can be scaled down

The Bad

Some optimization may be required to see if the primers are compatible with the shorter PCR program

The Bottom Line

The KAPA PCR mix is great for the large-scale genotyping PCR reactions done in our lab.

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