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pcr purification


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#1 siddharthsameer

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Posted 01 August 2012 - 09:39 AM

Hello
the concentration of my PCR product before purification was 1000microgram/ml and after the purification of my PCR product it has come down to 33.28 microgram/ ml.. what does it indicate.. is everyhing going fine or there is some problem in my pcr product..
kindly tell me....I am a big confusion. kindly help me..

#2 phage434

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Posted 01 August 2012 - 01:11 PM

Your unpurified PCR product has high concentrations of primers and dNTPs, which count as nucleic acid to the spectrometer. When you purify it, you get only the longer fragments, and see a huge reduction in apparent concentration. Measuring PCR products before purification is pretty meaningless.




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