Can DH5α be the host cell of the PCR-XL-TOPO from Invitrogen? - Help me (Apr/12/2006 )
Hi every one:
I wander whether DH5α can act as the Host of the PCR-XL-TOPO cloning vector from Invitrogen.
For the one shot Topo10 chemically competent cells provided in the Topo XL cloning Kit has expired.
Any suggestion is appreciated!
Thanks in advance!
Any one has the experiences on this?
I actually used DH5alpha cells (Invitrogen-the ones with subcloning efficiency. Cat. No. 18265-017) AND DH5alpha cells I had made chemically competent with the TOPO cloning kit from Invitrogen. And it worked well! As long as the cells have a good efficiency.
yes, i've done this on a production scale. works very well...
I would first test the Transformation efficiency of the Top 10 Chemically Competent cells with the test plasmid sent with the cells. If the cells still have a high transformation efficiency the Top 10 cells would still be ok to use. As long as the cells have been stored at -80C without freeze thawing the cells should be viable well after the expiration date (6 months).