PCR using random primer (hexamer)

Wilson netson at hotmail.com
Wed Apr 21 18:47:18 EST 1999


Dear all,
I have synthesized first strand cDNA molecules with different size by random
primer (hexamer) from RNA.  The kit that I used was SUPERSCRIPT
preamplification first strand cDNA.  Then I would like to synthesize second
strand cDNA molecules for cloning purpose.  My boss told me to use PCR with
random primer (hexamer again) for 2nd cDNA synthesis.  I would like to know
in detail about the PCR condition (no of cycle and annealing, denature,
extension temperature), amount of cDNA and primer should be used in the PCR
reaction.

Thank you very much

Wilson





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