sticky end to blunt end? - cloning (Aug/22/2007 )

Hello

I am going to combine a vector and insert which have no restriction enzymes sites in common.

Therefore I am thinking of cutting with sticky end cutters.
And then converting them to blunt ends (if it is possible??).
followed by ligation and transformation.

Has anyone tried the above method? I would like to see a protocol.

hallo matras,

there was a discussion in the past, which was about sticky-blunt end ligation

http://www.protocol-online.org/forums/inde...showtopic=28617

One can do that quite easily. You need the protocol for using klenow (to blunt the ends) which can be found in NEB's site.

we do it routinely.......no need to be scared........
but the blunt end ligation is difficult than sticky end ligation
to try to keep one end sticky