DDRT-PCR

yjiang yjiang at PUBLIC.EAST.CN.NET
Wed Mar 25 09:00:17 EST 1998


Dear Sir:

I've adopted DDRT-PCR for more than half an year in order to clone a
fertility-relative gene in Oryza sative. But I'm encountered a lot of
troubles. And I wonder if you can give me some good advice or help?
Could you give me some information about the different efficiency of direct
integration of a-32P and the terminal labelling of the anchored primers by
use of g-32P ? I've used the g-32P labelled anchored primers in the PCR and
got a greater resolving power, but at the same time, the false positive
frequency is greater. Is this caused by inappropriate ratio of anchored
primer and random primer? I wonder why researchers did not adopt the
terminal labelling of anchored primers ? Is it necessary to digest the RT
products in order to eliminate the high background caused by RNA? 
I'm now confused by my experimental results and eagerly looking forward to
your kindly help. Thank you very much!
 

Sincerely yours 
             Kangjian

                                                                98.March 23




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