PCR cloning

ellenmq at ucs.indiana.edu
Sun Jul 2 21:51:18 EST 1995


Junhyong (junhyong_kim at quickmail.cis.yale.edu) wrote:
: Hi,

: I heard somewhere that there are now other commercial T-tailed vectors
: available for PCR product cloning than TA-cloning kit from Invitrogen.
: Has anybody tried others? Is it cheaper?

: We tried making some but the results are rather variable...

We've used Novagen's T7-Blue TA cloning vector with good success. You
don't need the "kit" just buy the vector which makes it less expensive.
They recommend using  NovaBlue E.coli which has the same genotype as 
Stratagene's XL1-Blue cells. It's not necessary to use that particular 
E.coli strain, in fact we used a regular cloning strain of E.coli called 
DH11SF' available commercially from Gibco/BRL Life technologies. 
Alternatively standard DH5alpha cells should also be fine for propagation.  
You can check out Novagen's www site at this address: http://www.novagen.com/ 
for more information on this vector if you have a Web browser like Netscape 
available to you; they have both their catalog, vector sequences and 
plasmid maps available.

-Ellen Quardokus, Indiana University, Dept. of Biology




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