use of klenow to clone PCR products

GWDGV1::KCHOWDH kchowdh at gwdgv1.dnet.gwdg.de
Thu Nov 11 04:55:55 EST 1993


Hi Brian,
I routinely use Klenow to remmove the 3`overhang. All I do is to add the
enzyme directly to the restriction digest without any dNTP. In absence of
any nucleotide ( incubate 10 min at RT) the enzyme removes the 3`overhang
and may be little bit more. Then I add the dNTPs and incubate further 15
min to obtain a blunt end.
Cheers, Kamal.
Kamal Chowdhury
Max Planck Institute of Biophysical Chemistry
Am Fassberg
37077 Goettingen
Germany
Tel. 551-201507
Fax. 551-201504



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