Largest-Scale Ligation (Help)

Michael Benedik bchs1b at Elroy.UH.EDU
Sun Oct 25 22:17:06 EST 1992


In article <1992Oct24.000148.17185 at u.washington.edu>, pnj at stein.u.washington.edu (JnP) writes:
>Hi, do people know what the largest ligation could be (both
>theoretically and practically)?  As far as I know, those that 
>have been used for library constructions go up to a complexity 
>limit of 10^7.  Now I am wondering whether there is actually a 
>practical limit to the number of different sequences one can 
>handle in a single ligation reaction.  Also, I would like to 
>know the linear range (of insert/vector concentration) within 
>which one could get to increase the complexity.  (I have a 
>pool of degeneracy of 10^9 that I wish to handle, but I don't 
>think I will get to do 100 ligations or such.) 
>
>All opinions, experience, and advice are highly appreciated. 
>
>Many thanks!
>
>-Jing Huang
>

One obvious limiting factor to the complexity of ligations is 
transformation (packaging) or whatever you use to get it back
into cells. Although with super DNA and lots of tricks you should
be able to get 10x9, practically I have never heard of gettting
more than 10x7 after a ligation. Therefore I suspect this is the
true limiting factor.

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 Michael Benedik				INTERNET: Benedik at uh.edu
 Dept. of Biochemical & Biophysical Sciences	
 University of Houston				BITNET: Benedik at uhou
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