cloning systems - pGEM-T easy vector system versus TOPO TA cloning? (Oct/11/2005 )
Good day everyone,
I find myself in need of ordering a cloning kit and am not sure which to purchase. Which one do you recommend? Which is easier/more time efficient to use? Which one will be more successful with cloning an insert of about 2kb, and give a substantial number of white/blue colonies for selection?
Thanks for your anticipated input.
Gene Ome
topota cloning kit
I've had extraordinary good luck with the TOPO TA kits -- I honestly can't recall it ever failing...
I find myself in need of ordering a cloning kit and am not sure which to purchase. Which one do you recommend? Which is easier/more time efficient to use? Which one will be more successful with cloning an insert of about 2kb, and give a substantial number of white/blue colonies for selection?
Thanks for your anticipated input.
Gene Ome
I also have had good luck with TOPO TA kits, but I also have had equally good luck with pGEM. The primary advantage of pGEM is that it is cheaper.
I go from Accuprime Pfx (invitrogen) PCR amplified product to pCR-TOPO II BLUNT vector (invitrogen) every week with no problems. Of course, I'm biased because we get a 20% discount off of invitrogen products at Salk.
For taq polymerase (hopefully proof-reading), use their TA cloning vector.
One thing I like about this vector is its high efficiency, ccdb for removing positive background, and the close proximity of the M13 for and M13 rev sites to the cloning site.
-Matt