Would not the best way of ensuring or controlling for amplification from genomic
DNA be to use primers to the introns? Or perhaps PCR across a small intron so you
can see a difference in size in the PCR fragments of cDNA and genomic.
Best wishes, Martin Slade
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>To: biochrom at net.bio.net>From: JOHNSON YIP <n1607693 at sparrow.qut.edu.au>
>Subject: PCR primers
>Date: Wed, 9 Apr 1997 07:57:33 +1000
>>Hi there,
>> I've got a question here, and wish someone can help me....
>>When you design PCR primers to amplify from the mRNA, you need to consider
>some factors like GC content, Tm, internal 2ndary structure, 3'-end
>complimentarity, etc.. What other factors you need to consider if you are
>designing primers to amplify from the gene and not the mRNA??
>> Also if you want yuor primer to work only on cDNA, and want to detect
>whether there's any contaminate from the genomic DNA, how can you achieve
>this? (Southern blotting??)
>>Thanks
> My address:- n1607693 at sparrow.qut.edu.au>>>>---------------------------------------
Martin Slade,
School of Biological Sciences,
Macquarie University,
NSW 2109,
Australia
FAX (61 2) 9850 8174
Phone(61 2) 9850 8210
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