PCR-buffer development

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Topic: PCR-buffer development

Administrator
Administrator

posted 02-21-2001 01:41 PM

Our lab is involved in the screening for human mutations and the techniques involved in these procedures are dependant on PCR. I have therefore been trying to design a buffer that would work with most of our PCR reactions. Could anybody who might have gone through such trouble provide the insight on additives that worked for them either single or in combiantion.

Thanks,

Dumisani, University of Cape Town, SA

Posted by Gamad

dg@anat.uct.ac.za

IP: 160.45.191.21

JohnMoody
Member

posted 02-26-2001 10:58 AM

We went through a similar situation in our lab a while ago to look a polymorphic markers in the mouse. After trying many makes of TAQ and various alterations to the standard buffers we hit upon the then new TAQ from Qiagen. The TAQ in combination with their 'Q Solution' gave us better product yield, tighter bands and much reduced secondary banding which together enables us to routinely resolve 4 bp differences on ordinary agarose and 2 bp differences on acryamide or small fragment agarose. Why not give it a shot as it might work for you and save having to reinvent the wheel !!.

Hope this helps and best of luck.

IP: 194.129.135.161

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