TnT® Quick Coupled Transcription/Translation System

The TnT® Quick Systems are 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 p...

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The TnT® Quick Systems are 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.

Features - Benefits

• Obtain Data Faster: Functional protein is expressed in only one hour, not days as with cell-based expression systems.
• Multiple Applications with One System: Use expressed protein for the characterization of protein:protein interaction, protein:nucleic acid interaction, protein modification and more.
• Consistent, Reliable Results: This mammalian-based system expresses soluble, functional proteins that are post-translationally modified, unlike E. coli-based systems.
• Fewer Steps: Expressed proteins can be used directly after expression; no requirement for additional purification.
• Flexible Systems Available: TnT® Systems for linear, circular or PCR templates are available.

Applications

• 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).

For more information about cell-free protein expression, see the Protein Expression chapter of the Protocols & Applications Guide.

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