Tiny amount mRNA

[CK Wen]

ynishiwatari at UCDAVIS.EDU (Yutaka Ishiwatari) wrote:
>Dear Net-users,
>I am planning to make cDNA library from tiny amount of mRNA, less then 10 ng. Does 
>anybody have any experience for ampification of small amount of mRNA? Or does anybody 
>know good protocol for that kind of experiment? In case It's necessary, I can try to increase 
>the amount of mRNA little bit more up to 30 ng. But that's the maximam I can prepare. 
>
>Any idea would be very wellcome.
>Thanks in advance, and good luck!
>
>Yutaka

Dear Yutaka:

  I have never had this problem.  Could you make your first strand cDNA
with linker-oligo(dT)18 and TdT your first strand cDNA with oligo(dG).
Second strand cDNA could be synthesized by using linker'-oligo(dC). 
Thereafter, you may amplify your cDNA using these two linker sequences.

Chi-Kuang










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