Edited by Nayeem991, 10 July 2012 - 07:53 PM.
cloning without phosphorylation
Posted 10 July 2012 - 07:52 PM
Posted 10 July 2012 - 10:41 PM
Posted 11 July 2012 - 03:41 AM
Otherwise ask other lab(s) if they can give you a bit for the moment...
Posted 11 July 2012 - 11:47 AM
Be prepared to sort through large numbers of vector only colonies.
Posted 11 July 2012 - 08:59 PM
Posted 11 July 2012 - 11:28 PM
Posted 12 July 2012 - 03:03 AM
I don't know how it happened. but there is a minimal chance to happen this because the ligase buffer contains ATP and there might happen some spontaneous reaction to phosphorylate the insert. Its just chance. I think its not possible to get desired colony every time you perform the reaction without phosphorylating your insert. I tried several times and got it...
wow great news, but how is this possible when there is no phosphate group? I'm confused!
Posted 12 July 2012 - 04:21 AM
For me it also worked well but I only had to clone a resistence gene into a plasmid...