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From: werner@aecom.YU.EDU (Craig Werner)
Newsgroups: sci.bio
Subject: Re: Life Classification ...further comments
Message-ID: <1117@aecom.YU.EDU>
Date: Sat, 6-Jun-87 22:32:26 EDT
Article-I.D.: aecom.1117
Posted: Sat Jun  6 22:32:26 1987
Date-Received: Thu, 11-Jun-87 03:01:57 EDT
References: <9543@duke.cs.duke.edu> <1125@ius2.cs.cmu.edu> <701@edge.UUCP> <1302@sigi.Colorado.EDU>
Organization: Albert Einstein Coll. of Med., NY
Lines: 23
Keywords: Hanahan Transformation

In article <1302@sigi.Colorado.EDU>, pell@boulder.Colorado.EDU (Anthony Pelletier) writes:
> Even Doug admits that the protocol works that much better only on the
> DH cell lines.  For reasons that have never been clear to me, he does get
> the protocol to work on many other cell lines better than I (or anyone I know)
> have been able.  Something in his body temperature or "karma" I guess.
> It is well established that HB101 will not stand up to the Hanahan proceedure.
> It even says so in the "Joy of Cloning."

	I have gotten Hanahan transformation to work with HB101, as well
as JM101.  JM83 failed miserably.  I now use DH5alpha, as it has the
lacZ complementation of the JM series, but comes hyped.
	On my best day,  I got about 20% of Hanahan's maximum yield
and that was fresh.  Normally, I don't care, and use it frozen, and I
get 5-10% of theoretical. (Frozen also omits DTT)

	Nowadays I clone into phage.  It's much better for my purposes,
and it is much more efficient by anybody's standards, which is 10-100 fold.

-- 
			      Craig Werner (MD/PhD '91)
				!philabs!aecom!werner
              (1935-14E Eastchester Rd., Bronx NY 10461, 212-931-2517)
                      "If I don't see you soon, I'll see you later."