exclusion threshold for PCR purification
khatipovNO-SPAM at NO-SPAMuchicago.edu
Mon Dec 16 20:42:05 EST 2002
Here is example of such an abuse. From what I remember e.g. about the Qiagen
gel extraction kits, one is supposed to add isopropanol to the melted gel in
order to increase yield of fragments shorter than 500bp. Thus, omitting
IP-OH would do the job for Denis. The question is whether there is a 96-well
adaptation of the method, which I don't know.
"Wolfgang Schechinger" <hubahopp at gmx.de> wrote in message
news:220.127.116.11.20021216232927.007c8e90 at pop.gmx.net...
> You also might check the catalogues and tech specs from quiagen and
> macherey nagel. I think, they contain some information on different
> fragment exclusion sizes of different kits. So one might deliberately
> them for this purpose. I also think that binding and elution conditions
> (decrease of salt and ethanol) should cause loss of short fragments. Maybe
> their tech people can give you some details.
> At 10:10 PM 2002/12/16 -0000, Deanne Bell wrote:
> >:-----Original Message-----
> >:From: denis dupuy [mailto:denis_dupuy at dfci.harvard.edu]
> >:Does anybody know a way to modify this cuttoff so that I could retrieve
> >:only bigger fragments and get rid of my contaminant 100bp-species?
> >:Thank you in advance
> >This idea is attacking the problem form a different vantage point -
> >Have you tried doing a dilution / optimization / titration with different
> primer concentrations?
> >I am thinking that the problem may disappear by decreasing your primer
> >Deanne Bell
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