PCR cloning

[Edward Wang]

ellenmq at ucs.indiana.edu wrote:
: 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

Promega makes a kit that is significantly cheaper then Invitrogen.  I 
believe the kit is called p-Gem system
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Edward H. Wang                 *
Staff Resch. Assc.             *
Dept. Internal Med. -Nephrology*
UC Davis                       *
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