cloning genomic DNA fragments
pzheng at VAXA.WEEG.UIOWA.EDU
pzheng at VAXA.WEEG.UIOWA.EDU
Tue Jul 19 00:34:08 EST 1994
Dear bionetters:
I am desperate, would someone out there help me?
I had a lot of troubles in sucloing DNA fragments into pBluescript. By
using a
cDNA probe to screen a EMBL 3 genomic library, I was able to get a
positive clone.
By using different restriction enzymes, I was also able to generate
DNA fragments
of different sizes that can hybridize to the cDNA probe. However, the
troubles
started from there. When I tried to clone those fragments[blunt-
ended as well as
sticky-ended] none of them could be ligated into Bluescript[no
recombinants on
plants. And I believed the DNA fragment isolation and ligation
precedures had no
problems. Anyone ever had similar problems? Also, where can I get
primers
flanking the EMBL 3 cloning site? Thank you for your help! John
Zheng.
PZheng at vaxa.weeg.uiowa.edu
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