The RTS Linear Template Kit Plus accelerates the generation of multiple templates for parallel protein expression. Constructs are suitable for expression in E. coli and insect cell-free lysates. Due to compatible restriction sites in the adapter primers, products can be easily cloned into pIX3.0 vectors for large-scale in vitro expression.

The kit contains primers providing regulatory elements required for optimal transcription and translation in cell-free expression systems. Specially designed 5'-untranslated regions reduce the formation of secondary structure in the translation initiation region, one of the most common causes of low expression rates. A His-tag or Strep-tag II can be added to either terminus, greatly simplifying protein purification and detection.

A signal sequence enables expression of functional proteins containing disulfide-bonds or membrane proteins in RTS 100 Insect Membrane or Disulfide kits. A signal peptide can be introduced using the N-terminal primer, which contains a melittin sequence. This enables protein translocation into microsomal vesicles derived from insect cell endoplasmic reticulum.

(former FastLane product codes: RiNA C-1223-20)

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.

When used together with the RTS 100 E. coli HY Kits, the fast procedure enables determination of the optimal template structure within a single workday. The linear templates can also be used to synthesize posttranslationally modified eukaryotic proteins or membrane proteins using the RTS 100 Insect Membrane Kit. Expression screening of disulfide-bonded proteins, for example, antibody fragments, can be performed in 1-2 workdays when the kits are used in combination with RTS 100 E. coli Fab or RTS 100 Insect Disulfide kits. The system is highly suited for high throughput approaches.

Procedure
In the first PCR step, template DNA is amplified using gene-specific sense and antisense primers. In the second PCR step, DNA generated in the first PCR is the template and amplified using adapter sense and antisense primers.

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

Product manual

pQE2-EF-Ts sequence

Other documentation