Taq Polymerase for Mouse Genotyping

April 10, 2018

University of Rochester Medical Center
Microbiology & Immunology
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Qiagen

Product Name:

Taq DNA polymerase

Catalog Number:

201205

I manage a variety of transgenic and knock-in mouse strains and I have succeeded in genotyping using Qiagen Taq polymerase without even one case of failure.

Experimental Design and Results Summary

Application

PCR, genotyping

Starting Material

Mouse biopsy (tail fragment)

Protocol Overview

Any generally accepted protocol of PCR for genotyping with this Taq polymerase works. Here is an example of genotyping to determine mouse CD11a knock-out (one result was attached). Primer 1 (common F): GCAGCCCTGAGAAAGTTCC, Primer 2 (WT reverse): CTTCTGGTTTCCCCTGTCAC, Primer 3 (Mut reverse): CTCGTGCTTTACGGTATCGC, Reaction mixture (30 ul): 10x buffer 3 ul, 10mM dNTP: 0.6 ul, Primers: 0.2 uM each, Template: 1 ul of tail digested by proteinase K, Taq polymerase: 0.25 ul, water: rest of volume. PCR program: 95C 3min 1 cycle / 95C 15sec - 53C 20sec - 72C 20sec 35cycles / 72C 1 min 1 cycle.

Tips

This Taq polymerase works greatly once template is prepared appropriately

Results Summary

Multiplex PCR with this Taq polymerase discriminated WT, Het, and Homozygous mice nearly completely. In the attached figure, #1,8,9 were homozygous, #2,3,4,7 were heterozygous, and others were WT (Left-most lane contained DNA standards)

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Performance is high and price is affordable. And it is also very convenient because this PCR product is directly loaded on a gel without addition of loading dye/buffer.

The Bad

Nothing

The Bottom Line

Qiagen Taq polymerase was excellent for genotyping in almost any cases I have experienced.

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