Help on nested deletion system

[bio.tamu.edu]

In article <460lpo$p9h at oac4.hsc.uth.tmc.edu>, sa95078 at odin.mda.uth.tmc.edu
(sa95078) wrote:

> Hi, can anyone give me some suggestion for a reliable unidirectional
> nested deletion system.
> 
> Thanks in advanced.
> 
> James

Hi there,

I have had success using a DNAse deletion strategy which can be found as
follows:

Lin, Lei and Wilcox (1985) 
An improved DNA sequencing strategy
Analytical Biochemistry 147: 114-119

The only modification I made to the procedure was to replace all the DNA
precipitation steps with Promega's magic (now wizard) cleanups.  This
increased my yield of colonies about 1000-fold.  I'm sure other DNA clean
up columns would also work (no affiliation with Promega etc).

Hope this helps

Kim Snowden
ksnowden at bio.tamu.edu