Amplification of Plasmid Library

Mr. M.J. Lush mlush at hgmp.mrc.ac.uk
Mon Dec 7 09:47:28 EST 1998


	I'm making a cDNA expression library in a plasmid vector.  
I plan to divide the library into 125 pools of about 5000 clones each
during the amplification stage (this is because I'm screening via a 
bio assay).

	Methods in Enzymology (Vol152 1987 pp407-415) suggests that 
the library should be grown in soft agar on LB agar plates to
minimize differential growth between colonys.

	Whats the simplest method of seperating the bugs from the agar
(MiE suggests something fiddly involving converting the agar into a paste
by squerting through a 21 gague hypodermic needle and spinning through
a Sephadex column).

	Are there any better methods lurking out there?

TIA^6!




-- 

Michael
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
NPC rights activist           |      Nameless Abominations are people too.



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