The RTS Wheat Germ LinTempGen Set, His6-tag can be used to rapidly generate linear expression constructs that are ready for use with RTS Wheat Germ 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 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 the RTS pIVEX Wheat Germ vector and used with RTS 100/500 Wheat Germ CECF kits.

(former product codes: RiNA N-3540-96, 5PRIME 2402000)

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.

In order to generate linear expression constructs, two PCRs must be performed. In the first reaction, gene-specific primers are used to add overlap regions to the sequence of the target gene. The product of the first PCR is mixed with two flanking primers and DNA fragments coding for the T7 promotor, eukaryotic 5´- and 3´- regulatory elements and a His6-tag sequence (C-terminal or N-terminal). In the second overlap extension PCR, the product of the first PCR anneals with the added DNA fragments and the 5´and 3´ ends are extended. This linear expression construct is then amplified via the flanking primers.

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

Product manual

Other documentation