Path: utzoo!utgpu!watserv1!watmath!uunet!bionet!NRCCAD.NRC.CA!NUM208JN From: NUM208JN@NRCCAD.NRC.CA (JOHN NASH) Newsgroups: bionet.molbio.methds-reagnts Subject: Re: Ligations Message-ID: <9009082352.AA08801@genbank.bio.net> Date: 8 Sep 90 01:40:00 GMT Sender: daemon@genbank.bio.net Lines: 33 Hi there, MN>I have been suddenly having the utmost frustrating experiences with ligations May I ask a few questions?? I need more information. 1. What is the replicon of the plasmid? Will it replicate in the host you are transforming? What is the host in question? Which method of transformation are you attempting? 2. Can you transform uncut plasmid? What is your selection for transformants? Does the selection require "expression" before plating? 3. What amounts of DNA are you trying to ligate and transform? What restriction enzyme are you using? Can you detect ligation on a mini-gel? These questions may seem a bit trivial, but to a reader at the other end of the message (like me), it makes a difference between speculation and a more educated guess (is there a difference ;-) ?? ) cheers, John, -------------------------------------------------------------- John H.E. Nash Institute for Biological Sciences, National Research Council of Canada, Ottawa, Canada K1A 0R6. Phone: (613) 990-0990 Fax: (613) 952-9092. ==============================================================