The RTS 100 E. coli LinTempGen Set is designed to rapidly generate linear expression constructs that are ready for use with RTS 100 E. coli HY, Fab and insect kits. PCR generated linear templates allow fast protein expression from wild type or modified DNA sequences. Use PCR-based template generation to:

• Add epitope-tag sequences for antibody detection
• Introduce mutations
• Change codon usage
• Generate truncated proteins

Using linear DNA templates for protein expression avoids the time-consuming step of cloning into expression plasmids and can be used for initial, rapid screening for expression. Gene-specific primers can be used for RTS Wheat Germ and E. coli LinTempGen sets enabling easy transfer of products from any expression system to both the RTS Wheat Germ and E. coli platforms. For expression scale-up, the linear templates can be cloned into RTS pIVEX E. coli vectors and used with RTS 100/500/9000 E. coli CECF kits.

(former product codes: RiNA N-3510-96, 5PRIME 2401000)

Quality Control

in vitro transcription/translation protein expression products are manufactured using quality chemicals and materials that meet our high standards. All product components are subjected to rigorous quality assurance testing process:

• Component testing: each component is tested to ensure the composition and quality meet stated specifications.
• Performance testing: each product is tested to ensure it meets the stated performance specification.

High-Yield expression in the RTS requires cDNA templates that carry regulatory elements (T7-promoter, RBS, T7-terminator) with appropriate spacing. These elements can be added to a target gene either by standard cloning into pIVEX vectors (directly resulting in a circular expression template) or by overlap extension PCR using RTS Linear Template Generation Sets (resulting in a linear template). Linear templates show limited stability and can therefore only be used in RTS 100 E. coli HY short-run batch reactions, whereas the longer-lasting RTS 500 and RTS 9000 reactions require circular expression plasmids. If linear templates are used for RTS 100 reactions as a first step, they can be easily circularized later using standard blunt-end or TA-cloning techniques.

Use Linear Template Generation Sets

• to quickly generate mutational variants of your gene.
• to compare different clones or expression conditions in small scale RTS 100 reactions.

Use pIVEX vectors

• to express genes that do not need optimization.
• for medium or large-scale expression.
• as an alternative to templates generated by PCR.

For detailed protocol, please, refer to the product manual (Related Documents).

Product manual

Other documentation