E. coli S30 Extract System for Linear Templates Technical Bulletin

Instructions for Use of Product(s)
L1030

Literature # TB102

The E. coli S30 Extract System for Linear Templates allows transcription/translation of linear DNA templates. The system is used with linear DNA containing a prokaryotic E. coli-like promoter and sequence for a ribosomal binding site. In vitro-generated RNA from DNA templates lacking an E. coli promoter also may be used, but protein yields will be decreased to 1–10% of that produced from linear templates. The S30 extract is prepared from an E. coli B strain (SL119) deficient in OmpT endoproteinase, lon protease and exonuclease V (recBCD) activity, resulting in greater stability of expressed proteins. The recD mutation of the SL119 strain produces a more active S30 Extract for Linear DNA than nuclease-deficient, recBC-derived S30 extracts.

An easy-to-perform, non-radioactive positive control reaction using the Luciferase Assay Reagent provided allows detection of luciferase gene expression. The control reaction produces high light output for several minutes, allowing researchers to choose from several methods of detection, including simple visual observation of luminescence.

Summary of Changes

At the 6/15 revision of this document, expired patent statements were removed and the document design was updated.

Printed in USA. Revised 6/15.