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What fold change on the TaqMan = DNA duplication?

TaqMan qPCR rtPCR quantitation DNA

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#1 Gingeneticist

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Posted 05 December 2013 - 03:13 AM

Hi All, 

 

I have a DNA sample with a known duplication. I am trying to confirm the duplication via TaqMan assay with SYBR green.

I have a primer set within the duplication and one in normal sequence upstream - similar product sizes/Tms

So I expected that I could run my sample with the duplication and some control sampes for both primer sets and do the Ct>dCt>ddCt>fold change calculation to see that there are 2 copies of the normal primer set in my test and controls and 3 copies of the duplication primer in my test versus 2 copies in my controls

 

What I have got so far is a fold change of 7 for the duplication primer in the test subject

Equal amplification for test and controls in normal primer

 

What does this mean - is it saying I have 5 copies of the sequence I think I have 3 copies of? Does the fold change you calculate relate exactly to number of copies or is it on some kind of scale? I am confused

 

Massively appreciate any help, haven't found anything about this online so far!

 







Also tagged with one or more of these keywords: TaqMan, qPCR, rtPCR, quantitation, DNA

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