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PCR internal control - how does it work? (Jan/17/2006 )

ohmy.gif Confused, person here. I've been given the task of incorporating a PCR internal control into our assay, specifically, GAPDH. Basic question here: How does the internal control work? Is there a website that can explain this in layman's terms? Please forgive my ignorance. Thank you in advance. smile.gif

-Lewis F-

The internal control is used to verify identical sample loading in each assay. When there are many separate PCR reactions that will eventually be comapared to each other on a gel, it helps to know (via an internal control or otherwise) that each reaction used the same amount of starting material.

Since you're planning to use GAPDH as a control, I'm assuming you intend to run RT-PCR. In this case the GAPDH RNA levels are maintained by the cell and can thus be used as a PCR control. One would expect to see no variation in the GAPDH amplicon.