Why I always get smear in the third step of TAIL-PCR?
Posted 10 November 2004 - 07:13 PM
Posted 12 November 2004 - 07:27 AM
It depends for which purpose you need to conduct the 3rd step, because I've read that for nice sequencing two steps are enough.
Moreover, I was wondering if YOU could help me.
I have being tried to recover flanking regions using TAIL-PCR and until now, only unexpected products showed up, meaning that sequences of the transformated plasmid outside of LB-RB region appeared. Another problem is getting no product at all (or I get multiple bands patterns, including negative controls). I tried to use different TAQ polymerase, but results remained the same. I was wondering if you have any ideas or if you encountered similar problems and perhaps have suggestions for me. Do you think that it is possible for all the plasmid to integrate into the fungal DNA?
I would be extremely thankful for any of your ideas or suggestion.
Posted 07 March 2005 - 01:54 AM
Posted 07 March 2005 - 01:58 AM