Home-Made TA cloning

Ted M. tedm at darkwing.uoregon.edu
Fri Jul 21 22:34:47 EST 1995


Martin is correct, using Xcm I on pDK101, a derivative of pGEM5z, I
belive, will give a TA cloning vector. The plasmid is available from ATCC
, in JM109 host, for about $50
                                                   Ted Michelini



> 
> Interestingly, there is an enzyme (Xcm I)..possibly from New England
> Biolabs.....
> 
> the digestion product from this is
> 
> .......xxx|xxx....
> .......xx|xxx.....
> 
> or something like that...basically you can construct a vector that contains
> this enzyme site and have it so that two T's are left overhanging upon
> digestion...this is the basis of some vectors such as pDK101...and probably
> those sold by the biotech companies...
> 
> If you request this vector on this newsgroup...there are still some people
> that will freely distribute it to you....I was sent some but cannot send it
> off since it is not originally mine...saves buying the digested T vector from
> promega, clonetech, stratagene etc...
> 
> ......ahh just found Xcm I....it is from New England Biolabs and the site is
> 
> ...CCANNNNN|NNNNTGG...
> ...GGTNNNN|NNNNNACC...
> 
> 
> i believe this method of T-vector construction was published in Biotechniques
> a year or twoo ago...
> 
> good luck
> 
> Martin
> 
> .....          Martin Leach                Email:leach at bu.edu 
>    _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
>    / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
>  _/  |-/__==/  80 E. Concord St. (L603)
> (BULLDOZER) \_ Boston MA 02118            "Not the old underpants 
>                USA                                on your
>                                              head.....WIBBLE"
> 
> My homepage: http://155.41.115.114/Leach.h
> tml
> Biotech companies on the net: http://155.41.115.114/jumper/biolcomm.html



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