re-amp of PCR -- problems

Shahram Mori smori at nmsu.edu
Thu Jan 12 00:53:10 EST 1995


zool201 at csc.canterbury.ac.nz wrote:
: To whom it may concern

: I'm trying to reamplify a very low concentrartion of primary PCR
: product and am ending up with smears every time.
: I don't want ot re-purify the primary product because of the very low 
: concentration.

: Any hints would be greatfully accepted.

: Andrew Holyoake 
: PhD student
: University of Canterbury
: Christchurch 
: New Zealand

: I am trying to reamplify very low concentration primary PCR

Andrew,
What is the specificity of your random 10mers? are you using this in
differential display? What about ligating something to the 5'end of your
DNA and using primer to that with combination to the 3' primer. Also
it MAY be that your cycles are too many and you are getting amplification
of other nonspecific products rather than the one you are looking for.
Good Luck,
Cheers.



--
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
88003





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