DD-PCR

Youhua Xie xie at assel.biochem.mpg.de
Wed Nov 6 05:25:36 EST 1996


wudc at NIC.BMI.AC.CN wrote:
: Dear everyone

: The ones who have experince doing differential display,I want your help!

: I used 0.2 micrograms total RNA to reverse transcription,and one-tenth
: of the cDNA was used in DDPCR,but I got very faint bands,I think the
: reason may be too little cDNA was used in the reaction.so I want to
: know,how much total RNA is used to do reverse-transciption ,and how much
: cDNA is used to do DDPCR in your reaction?
: If you have some tips or skills ,let me know.Thank you very much!
0.2 ug total RNA is enough for a DD-PCR. You may have to check the efficiency
of your cDNA synthesis. Alternatively, you can use P33 instead of S35 .

Good luck




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