XbaI diestion of PCR products

Wolfgang Schechinger hubahopp at gmx.de
Thu Jul 22 16:41:01 EST 2004


Dear Raymond, 

PvuII and XbaI have completely different recognition sites. What's your
reason to use it as reference for XbaI (except that it is an RE working in
simalr buffer conditions?)

Might be that your DNA material is heterocygous, so there it's likely that
polymorphism is present. Would explain everything :-)

However, you cite this link to pall. Does this mean that you have the same
findings (i.e. you get the digest with size exclusion columns, but only
partial with EtOH precipitation?)

Best reagrds,

Wolfgang



At 13:00 22.07.2004 +0100, you wrote:
>We are having problems digesting PCR-amplified DNA with XbaI. It's for RFLP
>testing and the site is well within the amplified fragment, not at the end.
>The digests are always partial, and cleaning the DNA on Qiagen only seems to
>give a marginal improvement. The fragment digests correctly with PvuII
>without any clean up.
>
>Information on the Pall website (http://www.pall.com/924_25460.asp - see end
>of article) alludes to the issue.
>
>Does anybody else have this problem with XbaI, and how do you work around
>it?
>
>Thanks,
>
>Raymond
>
>
>
Wolfgang Schechinger


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