Hot Start LA Taq DNA Polymerase For Long Range (5-10 Kb) PCR

February 21, 2017

Albert Einstein College of Medicine
Clinical Research
Pre-doctoral researcher

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

TaKaRa Bio USA, Inc.

Product Name:

LA Taq DNA Polymerase Hot Start Kit

Catalog Number:

RR042A

We use TaKaRa's LA Taq DNA polymerase hot start kit to amplify up to 10 kb of genomic DNA by conventional PCR. Our focus is the amplification of entire parasite genes, and our starting material is parasite gDNA isolated and purified from infected red blood cells of human cases. This amplification kit has low background amplification, is easy to use as reactions can be set up at room temperature, and it is powerful enough for our long range amplification needs.

Experimental Design and Results Summary

Application

Long range amplification by convential PCR

Starting Material

Purified gDNA

Protocol Overview

Prepare a master mix according to the manufacturer's guidelines (Taq, buffer, and dNTPs -- all included in the kit; primers -- not included, experiment specific). Depending on the concentration of your gDNA preparations, add template to each reaction vial. The thermal cycling protocol will be dependent on the composition of the DNA samples as well as the target of interest for a given protocol.

Tips

Prepare a master mix when dealing with multiple samples.

Results Summary

Successful amplification of the gene of interest can be determined by visual detection of the expected band size following gel electrophoresis. Expected products can be excised from the gel and purified for subsequent sequencing analysis.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Time efficient, simple protocol -- can set up reactions at room temperature, reliable for long range amplification needs, low background.

The Bad

Slightly expensive.

The Bottom Line

I highly recommend this product for long range amplification of genomic DNA. Anyone who has performed long-range PCR knows that it can be a pain, and this product makes a world of difference in quality and specificity of the amplification product(s).

Join the discussion