RTS Wheat Germ LinTempGen Set, His6-tag

One of the major advantages of cell-free compared to cell-based expression systems is the ability to use PCR-generated templates. Using the RTS Wheat Germ Linear Template Generation Set, His₆-tag, if primers are designed for the first (gene specific) of the two PCRs according to the guidelines provided in the pack insert, the resulting product can be used in a second PCR to produce a completely functional linear expression template for expression in either the RTS 100 E. coli HY Kit (using RTS E. coli Linear Template Generation Set, His₆-tag. The design of these kits enables easy transfer of products from any expression system to both the RTS Wheat Germ and E. coli platforms, as well as between the two RTS systems. 
At a later step in the RTS workflow, insertion of these products into an RTS pIVEX Wheat Germ vector may be required (e.g., to produce a template for expression scale-up to RTS 500 Wheat Germ CECF, or to ensure that a well-characterized (sequenced) and stabilized template is used in a series of subsequent experiments).

The RTS Wheat Germ product line combines a series of new technologies for efficient and optimized protein expression in eukaryotic cell lysates. It overcomes the limitations that often restrict the use of E. coli-based systems (cell-based or cell-free), especially with eukaryotic target proteins.

The RTS Wheat Germ Linear Template Generation Set, His₆-tag can be used to rapidly generate linear expression constructs by PCR, ready for use as DNA templates in RTS 100 Wheat Germ CEDF and RTS 500 Wheat Germ CECF protein expression reactions. The use of PCR instead of subcloning into vectors allows rapid protein expression from wild-type or modified DNA sequences. Use PCR methods to:

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

This method of generating 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.