A Robust Proofreading PCR Kit

Osmania University
Department of Biochemistry
Research Fellow

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Company:

Sigma-Aldrich

Product Name:

AccuTaq LA DNA Polymerase

Catalog Number:

D8045-125U

Molecular techniques like cloning require PCR amplicons without any mismatch in the amplicon sequence. Conventional PCR kits have a high rate of mismatch mutations because of lack of proofreading activity of the Taq. AccuTaq is a recombinant Taq with 3' proofreading activity, which of greatly helpful in the cloning process.

Experimental Design and Results Summary

Application

Cloning

Starting Material

cDNA/genomicDNA

Protocol Overview

This kit can be used by following manufacturer's instruction. Briefly, one unit of enzyme is used per reaction with an appropriate template, primers, dNTPs, and buffer.

Tips

Do not attempt to take more than one unit of enzyme as the enzyme is robust and has high fidelity.

Results Summary

Clear cut bands are observed with high accuracy

Additional Notes

None

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Summary

The Good

Very robust and efficient

The Bad

I did not face any problems.

The Bottom Line

I strongly recommend this kit for cloning of an interested gene.

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