cloning PCR products into vectors

Morris F. Manolson mm6y+ at andrew.cmu.edu
Wed Jul 7 15:14:09 EST 1993


Dear bio-netters,  I would like to take a survey about the method 
of choice for cloning PCR products into vectors.  Please vote for what
you beleive is the superior method.  I will tabulate and post the 
results in one week. 

Method 1) TA cloning
 
       2) filling in the overhang, and doing blunt-end ligations
 
       3) designing oligonucleotides with restriction sites on the 5' 
          ends

       4) others


For those of you doing TA cloning,  I would also like to know the source
of vector with the 3' T overhang.  I remember a post on materials and methods
a while back where someone constructed a plasmid with a tandom copy of one
restriction site which when cut left overhanging Ts...does
this ring a bell with anyone?  Thanks in advance for all votes and comments.
Morrie  


******************************************************************************
Morris F. Manolson                     Tel:  416-813-6662  (office)
Division of Cell Biology                     416-813-5594  (lab)
Hospital for Sick Children                   416-813-5028  (FAX)
88 Elm St., McMaster building        email:  Morrie at resunix.ri.sickkids.on.ca
Toronto, Ontario, Canada
M5G 1X8



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