pcr yield

Mr. C.S.Wilding osp086 at clss1.bangor.ac.uk
Tue Sep 20 07:33:06 EST 1994


I am attempting PCR on a 2kb fragment of scallop mtDNA using as template a
reasonably high purity solution of mtDNA. I got as much mtDNA as I could
from approx 5g muscle, dissolved in 50ul, put 1ul in 100ul dH2O and used 1ul
for PCR. This worked fine to start but now from some animals it doesn't- the
band is there but low yield. If I use 1ul of the undiluted DNA the band is
back up to good yield. Now I can understand that for some scallops I might
have got more DNA out- even up to say 10X more, but the PCR should be able
to account for this within a few cycles. It does seem like a "waste" of the
PCR to be putting what I believe is quite alot of DNA in the reaction.
I also get a low yield band from genomic DNA templates.
Has anyone any ideas why this should be?
Any hints on increasing yield- I have, I believe optimised with respect to
ann. temp., Mg2+.
Thanks, Craig.
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     * 	CRAIG WILDING,		       E-mail: OSP086 at BANGOR.AC.UK   *
     *	SCHOOL OF OCEAN SCIENCES,      Phone: (0)248 351151 ext.2899 *
     *	UNIVERSITY OF WALES (BANGOR),  Fax:(0)248 716367  	     *
     *	MENAI BRIDGE, 	           				     *
     *	GWYNEDD, LL59 5EY,					     *
     *	WALES, U.K.						     *
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