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Abstract for TNT® Quick Coupled
Transcription/Translation Systems
The TNT® Quick Coupled
Transcription/Translation Systems are convenient single-tube, coupled
transcription/translation reactions for eukaryotic in vitro translation. The
original TNT® Coupled Reticulocyte Lysate
Systems simplified the process and reduced the time required to obtain in vitro
translation results compared with standard rabbit reticulocyte lysate systems.
Standard rabbit reticulocyte systems commonly use RNA synthesized in vitro from
SP6, T3 or T7 RNA polymerase. The TNT® Quick
Coupled Transcription/Translation System further simplifies the process by
combining the RNA polymerase, nucleotides, salts and Recombinant RNasin®
Ribonuclease Inhibitor with the reticulocyte lysate to form a single TNT®
Quick Master Mix. For most gene constructs, the TNT®
Quick reaction produces significantly more protein (two- to sixfold) in a
60- to 90-minute reaction than a standard in vitro rabbit reticulocyte lysate
reaction using RNA templates. The TNT® Quick
Coupled Transcription/Translation System is available in two configurations for
transcription and translation of genes cloned downstream from either the T7 or
SP6 RNA polymerase promoters.
Included with the TNT® Quick System is a
luciferase-encoding control plasmid and Luciferase Assay Reagent, which can be
used in a non-radioactive assay for rapid (<30 seconds) detection of
functionally active luciferase protein. Starting with either circular plasmid
DNA or PCR-generated DNA, in vitro transcription/translation results (autoradiograms)
may be obtained easily in 5–6 hours.
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