Tnt sp6 quick coupled kit

Manufactured by Promega

The TNT SP6 Quick Coupled Transcription/Translation System is a cell-free expression kit that enables coupled in vitro transcription and translation of DNA templates. The kit provides a rapid and efficient method for the synthesis of proteins from DNA templates.

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Lab products found in correlation

Megaclear transcription clean up kit, by Thermo Fisher Scientific (2 mentions) Rabbit reticulocyte lysate system, by Promega (2 mentions) Mmessage mmachine sp6 kit, by Thermo Fisher Scientific (2 mentions) 35s methionine, by PerkinElmer (2 mentions) Sucrose, by MP Biomedicals (1 mentions)

Close spelling variants of this product

TNT kit (18 citations)

2 protocols using tnt sp6 quick coupled kit

In Vitro Protein Synthesis Protocol

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For in vitro translation of
³⁵S-radiolabeled precursor proteins, plasmids carrying a SP6
promoter-binding region followed by the gene of interest were used. Plasmids
were incubated with the TNT SP6 quick coupled kit (Promega), an in
vitro eukaryotic transcription/ translation system based on
rabbit reticulocytes, and [³⁵S] methionine (PerkinElmer) for 2 hr
at 30°C and 300 rpm.
For the in vitro translation of
[³⁵S]Por1 the Rabbit Reticulocyte Lysate System (Promega) was
used. First, mRNA was synthesized by using the mMESSAGE mMACHINE SP6 kit
(Invitrogen) and cleaned up by the MEGAclear Transcription Clean-Up kit
(Invitrogen). Radiolabeled proteins were translated by using the Rabbit
Reticulocyte Lysate System (Promega) for 2 hr at 30°C and 300
rpm.
After translation, 20 mM unlabeled methionine (Roth) and 0.3 M
sucrose (MP Biomedicals) were added. Lysate was snap frozen in liquid
nitrogen and stored at –80°C.

Weinhäupl K., Lindau C., Hessel A., Wang Y., Schütze C., Jores T., Melchionda L., Schönfisch B., Kalbacher H., Bersch B., Rapaport D., Brennich M., Lindorff-Larsen K., Wiedemann N, & Schanda P. (2018). Structural Basis of Membrane Protein Chaperoning through the Mitochondrial Intermembrane Space. Cell, 175(5), 1365-1379.e25.

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in vitro Translation of Radiolabeled Proteins

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For in vitro translation of ³⁵S-radiolabeled precursor proteins, plasmids carrying a SP6 promoter-binding region followed by the gene of interest were used. Plasmids were incubated with the TNT SP6 quick coupled kit (Promega), an in vitro eukaryotic transcription/translation system based on rabbit reticulocytes, and [³⁵S] methionine (PerkinElmer) for 2 hr at 30°C and 300 rpm.
For the in vitro translation of [³⁵S]Por1 the Rabbit Reticulocyte Lysate System (Promega) was used. First, mRNA was synthesized by using the mMESSAGE mMACHINE SP6 kit (Invitrogen) and cleaned up by the MEGAclear Transcription Clean-Up kit (Invitrogen). Radiolabeled proteins were translated by using the Rabbit Reticulocyte Lysate System (Promega) for 2 hr at 30°C and 300 rpm.
After translation, 20 mM unlabeled methionine (Roth) and 0.3 M sucrose (MP Biomedicals) were added. Lysate was snap frozen in liquid nitrogen and stored at −80°C.

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