Optimizing PCR reactions for gene amplification – Master Mix


If you are working in the geomicrobiology field or a geologist (or just curious) )who is interested in geomicro you will run into PCRs (Polymerase Chain Reaction) at some point in time. Basically the PCR lets you target a piece of DNA and then copies it a billion times or so. This means lots of it! Once you have this mess of segments you can then identify what it is. Pretty neat. The University of Utah has an excellent Virtual PCR program. I’d recommend checking it out.


So you collected some rocks that have buggies on them. You might want to know who the buggies are. Or maybe you are interested in seeing if they have genes that produce siderphores (compounds the scavenge metals). PCR can help with this (so can 454, which is another topic).

We use AmpliTAQ Gold because we can get it downstairs fairly cheaply. Not all TAQs are created equal. I am going to run an experiment later with different TAQs to compare each one.

Basic Master Mix recipe for AmpliTAQ gold

for 15 µL reaction

(volumes in µL)

H2O 9.57

10x PCR Buffer 1.5

100x BSA 0.125

Forward Primer 0.1

Reverse Primer 0.1

TAQ 0.15

MgCl2 1.5

There you go.

This entry was posted in Techniques and tagged , . Bookmark the permalink.

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out /  Change )

Google+ photo

You are commenting using your Google+ account. Log Out /  Change )

Twitter picture

You are commenting using your Twitter account. Log Out /  Change )

Facebook photo

You are commenting using your Facebook account. Log Out /  Change )


Connecting to %s