The TnT® Quick Coupled Transcription/Translation Systems provide convenient single-tube, coupled transcription/translation reactions for eukaryotic cell-free protein expression. These cell-free expression systems combine the RNA Polymerase, nucleotides, salts, amino acids and Recombinant RNasin® Ribonuclease Inhibitor with the reticulocyte lysate solution to form a single TnT® Quick Master Mix.
The TnT® Quick Coupled Transcription/Translation System is available in two configurations for the expression of genes cloned downstream from either the T7 or SP6 RNA polymerase promoters. To use these cell-free expression systems, 0.2–2.0μg of circular plasmid DNA containing a T7 or SP6 promoter, or a PCR-generated fragment containing a T7 promoter, is added to an aliquot of the TnT® Quick Master Mix and incubated in a 50μl reaction volume for 60–90 minutes at 30°C. The expression reaction produces significant quantities of protein for a variety of applications including GST pull-downs and gel shift assays.
These mammalian-based systems express soluble, functional proteins that are post-translationally modified, unlike E. coli-based systems. Expressed proteins can be used directly after expression; no additional protein purification needed.
For more information about cell-free protein expression, see the Protein Expression chapter of the Protocols & Applications Guide.
- GST pull-downs (view an animation).
- Gel shift assays (view an animation).
- Co-immunoprecipitation (view an animation).
- Characterization of protein modifications (view an animation).
- Protein:RNA interactions.
- Protein activity studies.
- Confirmation of gene products.
- Protein Truncation Test (PTT).