Very Easy and Efficient Taq Mastermix

January 28, 2019

Albert Einstein College of Medicine
Genetics Department
Ph.D. Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Promega

Product Name:

GoTaq® Green Master Mix

Catalog Number:

M7122

I needed a cheap and efficient PCR mix since I have to do a lot of PCR from bacteria colonies to check presence/absence of my target insert in the plasmid. I found this kit very easy to use; I didn't need to optimize any conditions, it works every time, and you can load it into the agarose gel right after the PCR reaction.

Experimental Design and Results Summary

Application

Bacteria colony PCR

Starting Material

Bacteria colonies, primers, and GoTaq mastermix

Protocol Overview

I pick the bacterial colonies with a tip and dissolve them in 10 µL of H2O each. I use 1 µL of the dissolved bacteria for each PCR reaction. Since primers are the same for all the colonies, I make a mastermix with the following components (quantities are for one single PCR tube): GoTaq 2X mastermix - 12.5 µL, Primer F (10 µM) - 2.5 µL, Primer R (10 µM) - 2.5 µL, H2O - 6.5 µL. Of this mix I distribute 24 µL into each PCR tube, which already contain 1 µL of dissolved bacteria. The thermocycler settings are the same ones indicated by Promega on the instructions.

Tips

When mixing the components in the mastermix, always calculate the quantities in excess (i.e. if you are doing 5 colony PCR, calculate for 5.5).

Results Summary

I always get good results, always have a DNA band on the agarose gel.

DOI or PMID #

N/A

Additional Notes

For obvious reasons, I wouldn't use this mix for sensitive applications like cloning or sequencing, since this Taq does not have a proofreading mechanism and could introduce mutations during DNA replication.

Image Gallery

Summary

The Good

Easy to use, works every time

The Bad

When pipetting the mix, I always get some reagents stuck in the tip, so you can lose small amounts of the mix.

The Bottom Line

Saves a lot of time when I have to do a high number of PCRs, since it has all the components necessary for PCR and a loading buffer.

Join the discussion