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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