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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:

  • Variety of DNA cloning experiments

  • Screening of nested deletion library

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.