Fast
Clone ID
Kit
(Catalog
# K004)
Description:
The
LAE-Fast
Clone ID Kit provides
a highly efficient screening method to identify
bacteria colonies transformed with your desired
DNA-fragment-containing plasmid. The
detection method is based on the change in
mobility in an agarose gel of a plasmid containing
your insert of interest versus an ˇ§emptyˇ¨
plasmid. In a typical cloning experiment, a
DNA fragment is ligated to a plasmid vector,
resulting in a larger plasmid. The ligation
mixture is then transformed into bacterial hosts
and propagated under selective conditions.
Usually a dozen bacterial colonies are picked and
cultured over night. Plasmid DNA is then
purified and analyzed by gel
electrophoresis. In some difficult cloning
experiments, DNA must be prepared from more
bacterial colonies for analysis. By using
the 5 minutes cloning screen kit, the time
consuming, tedious, and expensive DNA preparation
procedures can be eliminated. In one simple
step, the specially formulated Screen Buffer lyses
the bacteria. The cell lysate is then
separated by agarose gel electrophoresis.
The desired plasmids are then identified by their
electrophoresis mobility shift compared to a
vector control.
Applications:
Protocol:
1.
Pick cells from streaks and disperse the
cells or 10ml bacterial culture in 100 ml of SB (Screen Buffer). Leave at
room temperature for 5 minutes or until cell
lysed.
2.
Load 30-40 ml into wells of agarose gel. In a
separate well, load 0.5mg of parent plasmid DNA as a
control.
3.
Stain gel and view.
Kit Contents:
500 ml SB per
tube (containing glycerol, SDS, RNase A,
Bromophenol blue, and TAE); 10 tubes per
kit.
Shipping/Storage:
This kit is shipped at ambient
temperature and should be stored at
4oC.
Legal
consideration: FOR RESEARCH USE
ONLY. |