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cDNA concentration higher than original RNA


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4 replies to this topic

#1 Curtis

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Posted 19 January 2010 - 07:25 PM

Hi,

my RNA has 1.8 ng/ul concentration but the cDNA that I made from it has 1.2 ug/ul concentration. Even higher.

How is this possible when I used only 2 ul of RNA as tamplate in 20 ul reaction. I mean the cDNA concentration must be even lower, but now it is higher. How?

#2 molgen

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Posted 20 January 2010 - 01:48 AM

Did you take into consideration when doing the calculation that cDNA is both double stranded and a heteroduplex?

#3 Curtis

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Posted 20 January 2010 - 08:21 AM

I do know that, but during measurement with Nanodrop we consider cDNA as ssDNA. This is what the company told us. But still doesn't matter if we consider it as double strand or single strand, the cDNA conc. is still higher. is it always like that?

#4 bob1

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Posted 20 January 2010 - 03:12 PM

try putting some dNTPs through the spec/nanodrop...

#5 phucvn

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Posted 17 February 2010 - 11:44 PM

After purifying your cDNA, the resulting concentration would be much lower.




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