TA cloning: thanks!
dschnapp at molbiol.ox.ac.uk
Thu Jun 18 05:36:53 EST 1998
Thank you for all the advise on TA cloning. To summarize: apparently, the
Promega PGem kit is fine, but there might be some variation between kit
performance. Invitrogen Topo 2.1 kit is very good (but costly, I think).
It is best to chloroform-extract the PCR reaction at once to inactivate
Taq, but inactivation may be incomplete. A valuabe piece of advise was to
add more Taq to the product after storage and incubate at 70-72 deg. C for
10 min prior to cloning - I'm off to try that next!
>Dept. Paediatric Gastroenterology
>59 St. Bartholomews Close
>London EC1A 7BE
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