Cloning very large inserts - help

Paul N Hengen pnh at ncifcrf.gov
Thu Nov 2 13:40:55 EST 1995


| I'm having trouble cloning some large inserts into an 8 kb vector
| (Casper). I recently got a vector from Invitrogen and noticed that they
| provide a bacterial strain called "TOP10". Among other things, it includes
| a mutation called deo[R], which supposedly enhances plasmid
| transformation, particularly large constructs. 
|
| Has anyone had experience with TOP10 or the deo[R] mutation? Or, any
| advice on cloning large inserts? It may be vector specific, since I have
| had no trouble cloning fragments as large as 25kb in pBluescript and pGEM.
|
| Michael Myers
| Laboratory of Genetics
| The Rockefeller University
| 212-327-8233  (-7420 fax)
| myersm at rockvax.rockefeller.edu

You might try this method, if you are really having troubles.
I have not tried it myself though.

@article{Zhixing1995,
author = "Y. Zhixing
     and J.-L. Nahon",
title = "{DNA} gyrase improves {DNA} transformation of
{{\em E. coli}} cells with large recombinant plasmids",
journal = "Nucleic Acids Res.",
volume = "23",
number = "16",
pages = "3353-3354",
year = "1995"}

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