TOPO cloning problem
Posted 08 October 2010 - 04:59 AM
Posted 08 October 2010 - 06:44 AM
I think if your PCR works fine with your gene of intrest, Then its of no use to go to this step and directly you can go to pET cloning for expression.
and select the abc+ bactetria after Lactose induction than IPTG. As it will give bacteria a kind of another stress. ( One should use or i will recomend to use IPTG for high protein yield) thatīs not necessary for this type of functional assays .
Posted 08 October 2010 - 01:43 PM
Posted 08 October 2010 - 02:20 PM
Edited by bob1, 08 October 2010 - 02:20 PM.
Posted 11 October 2010 - 12:47 PM
IMO, I would do it right that do it again. Plus, TA cloning is very easy and fast. It may take you less than a 3 days to find the right clone. Later, you can subclone into the pET all you want.