Ligation: two basic questions
Posted 21 February 2013 - 03:02 AM
2. I have a mixture of two PCR products that I'd like to ligate in tandem, one connected to each other, into a vector. However, one of them is capable of ligating into the vector alone. Is this ligation even worth trying? Will there be 100% preference for the small single insert, or is there rather a good chance that I obtain what I wish?
Posted 21 February 2013 - 04:13 AM
2.ya nothing wrong in trying though the chances of getting what you wish is less, may be you might have to screen, screen and screen lot of colonies to get. Alternatively you can try a sequential ligation, like ligating the pcrs first and then adding it on to the ligation mix with vector alone, in this way you can increase the chances.
I always had an alternate hypothesis....