problem with simple cloning!

Denise Melanson dmmelans at unity.ncsu.edu
Sun Apr 13 06:14:43 EST 1997


I believe she is trying to kill her XmaI site by filling it in. In any
case, when we are trying to kill a site, we will digest with the enzyme of
interest, gel isolate the linearized band, fill in with klenow, then take
an aliquot of this reaction and religate but include the RE in the reaction
mixture, this ensures that any incomplete digestion products will not
religate improperly.  However, this is the strategy we use when we are
killing sites with oligos, have you thought of this yet? Design a very
short oligo to KILL the xmaI site, and then religate.  Good luck.




More information about the Methods mailing list