Hi,
How can I measure cDNA concentration? I used 1 microgram RNA to make cDNA. Does that mean I have still 1 microgram cDNA? Thanks.
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bob1
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Thelymitra pulchella
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Posted 17 January 2005 - 12:19 PM
Hi
You can measure cDNA concentration by any of the standard methods you use to measure nucleic acid concentration.
It is unsafe to assume that because you started with 1 ug RNA that you will end up with 1 ug cDNA. For this to happen you would need the reaction to go to completion, which never happens with enzyme mediated reactions. It would also assume that all of the RNA was primed by your oligo dT or gene specific primers, a situation that is unlikely to occur.
Anyway, Hope this is useful.
Bob
You can measure cDNA concentration by any of the standard methods you use to measure nucleic acid concentration.
It is unsafe to assume that because you started with 1 ug RNA that you will end up with 1 ug cDNA. For this to happen you would need the reaction to go to completion, which never happens with enzyme mediated reactions. It would also assume that all of the RNA was primed by your oligo dT or gene specific primers, a situation that is unlikely to occur.
Anyway, Hope this is useful.
Bob
