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PCR quantification


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#1 lukefreedman

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Posted 28 January 2009 - 06:50 AM

Hello all wise peoples. Im conducting end point PCRs using a number of primers pairs which im comparing to a baseline reading constant between conditions. Im told that using image J and some of its associated plugins i can quantify my pcr bands and and produce ratios relative to the baseline which can then be compared to each other. I had a search on the web but didnt realy manage to find any real instructions. Can anyone help, either by explaining the process itself or pointing me in the direction of someone/something that can? Any assistance would be realy appreciated, quantifying my results would add much weight to the project im doing.


Thanks.

Luke

#2 Unagi

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Posted 30 January 2009 - 02:50 PM

Hi Luke,

Sorry of not being of direct help, but have you considered using real time PCR, if a machine is available to you? Quantitation is much more accurate & reliable using rtPCR data than band intensity data (even if it is digitised).




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