three way ligation (two fragment and a vector) - (Sep/23/2005 )
Dear everyone,
Who knows how to do three-way-ligation(two fragments and a digested vector)? should I set up one ligation reaction by putting two fragments (3kb each) with the digested vector together into one tube and add T4 DNA ligase? Any protocol for this?
Thanks a lot,
-amstel-
QUOTE (amstel @ Sep 24 2005, 04:50 AM)
Dear everyone,
Who knows how to do three-way-ligation(two fragments and a digested vector)? should I set up one ligation reaction by putting two fragments (3kb each) with the digested vector together into one tube and add T4 DNA ligase? Any protocol for this?
Thanks a lot,
Who knows how to do three-way-ligation(two fragments and a digested vector)? should I set up one ligation reaction by putting two fragments (3kb each) with the digested vector together into one tube and add T4 DNA ligase? Any protocol for this?
Thanks a lot,
you may add the two fragments and the digested vector together provided there is no repeatation of retriction enymes sites; if not, self ligation of vector or repeatation of fragment ligation may happen. what is your restriction enzymes sites?
-alitan-