Ligation failed

Duncan Clark blackhole at abuse.plus.com
Fri May 5 08:41:54 EST 2006


Historians believe that in newspost 
<wqG6g.23191$vy1.10010 at news-server.bigpond.net.au> on Fri, 5 May 2006, 
newsnet customer <customer at newsnet.com> penned the following literary 
masterpiece:
>HERE ARE MY THOUGHTS:
>1. chemically competent cells are not suitable for vector >10kb - 
>HIGHLY PROBABLE

Does ccc expression vector transform and at what frequency? I would work 
on the assumption that if it is 10E7 per ug that you get back then 
assume 10E3-10-E4 maximum per ug of ligated material.

>2. ligase and/or ATP (in lig. buffer) is not functional/depleted - 
>HIGHLY PROBABLE

Simple to test. Cut something with a 4 cutter RE and see if it is 
ligatable using a gel to tell.

Duncan

-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.


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