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T7 kits

Manufactured by Thermo Fisher Scientific
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Sourced in United States

The T7 kits are a series of products designed for in vitro transcription and translation. The kits provide the necessary components, including the T7 RNA polymerase, buffer, and other reagents, to enable the synthesis of RNA from DNA templates containing the T7 promoter sequence. These kits are commonly used in research applications to produce RNA for a variety of purposes, such as mRNA synthesis, RNAi studies, and in vitro translation experiments.

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Lab products found in correlation

Tricaine methane sulphonate, by Merck Group (1 mentions) Rnaeasy kit, by Qiagen (1 mentions) T3 mmessage mmachine, by Thermo Fisher Scientific (1 mentions) Wheat germ extract system, by Promega (1 mentions) Ni nta purification kit, by Thermo Fisher Scientific (1 mentions) Sp6 mmessage mmachine, by Thermo Fisher Scientific (1 mentions) 35s met, by PerkinElmer (1 mentions)

Close spelling variants of this product

MegaScript T3 and T7 kits T7 or T3 mMESSAGE mMACHINE kits MMessage mMachine SP6 & T7 Transcription Kits T7 Transcription Kit MAXIscript T7 in vitro Transcription kit MEGAscript SP6 or T7 Kit MMESSAGE mMACHINE T7 Ultra Kit MMESSAGE mMACHINE SP6/T7 kit MEGAscript T7 Transcript Kit Transcript AidTM T7 High Yield Transcription kit

3 protocols using t7 kits

1

Xenopus Oocyte Electrophysiology Protocol

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cRNA was synthesized by in vitro transcription using message machine T7 kits (Ambion, Austin, TX, USA) and injected into stage V and VI oocytes which were surgically isolated from female Xenopus laevis (Nasco, Modesto, CA, USA) anesthetized with 0.17% tricaine methanesulphonate (Sigma, St. Louis, MO, USA). The injected oocytes were maintained in modified Barth`s solution which contained 88 mM NaCl, 1 mM KCl, 0.4 mM CaCl, 0.33 mM Ca(NO3)2, 1 mM MgSO4, 2.4 mM NaHCO3, 10 mM HEPES (pH 7.4), and 50 µg/mL gentamicin sulfate at 17°C. The currents were measured three to six days after injection.
Jiang Y., Piao J., Liu N., Hou J., Liu J, & Hu W. (2020). Effect of Ultrafine Powderization and Solid Dispersion Formation via Hot-Melt Extrusion on Antioxidant, Anti-Inflammatory, and the Human Kv1.3 Channel Inhibitory Activities of Angelica gigas Nakai. Bioinorganic Chemistry and Applications, 2020, 7846176.
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2

In vitro Synthesis and Cloning of Fluorescent Protein Plasmids

Cited 1 time
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To generate cRNAs, plasmids were linearized and in vitro transcribed using a mMessage mMachine T3 (Ambion #AM1348) and T7 kits (Ambion #AM1344), according to manufacturer’s protocol. The synthesized cRNAs were then purified using an RNAeasy kit (Qiagen #74104) and stored at -80°C. The pYX-EGFP plasmid was created by transferring T3-T7 cassette from pRNA-EGFP vector [66 (link)] into the pXY-Asc vector (NIH, Bethesda, MD, USA) using PCR cloning. The pYX-EYFP plasmid was created from pYX-EYFP plasmid by replacing coding sequence for EGFP by EYFP. AURKC coding sequence [31 (link)] was cloned by PCR into pYX-EYFP to create pYX-AURKC-EYFP plasmid. pIVT-AURKB/C-EGFP and pGEMHE-mEGFP-mCDK5RAP2 plasmids were described previously [22 (link), 31 (link)].
Blengini C.S., Ibrahimian P., Vaskovicova M., Drutovic D., Solc P, & Schindler K. (2021). Aurora kinase A is essential for meiosis in mouse oocytes. PLoS Genetics, 17(4), e1009327.
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3

In Vitro Reconstitution of GAIT Complex

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Capped, poly(A)-tailed luciferase (Luc) reporter upstream of the ceruloplasmin (Cp) 3’UTR GAIT element (Luc-Cp GAIT) and T7 gene 10 reporter transcripts were prepared using mMessage mMachine SP6 and T7 kits (Ambion), respectively (Jia et al., 2008 (link)). For in vitro reconstitution of GAIT complex, His-tagged S886A EPRS linker (permitting site-specific Ser999 phosphorylation) was phosphorylated by immunocomplexed Myc-S6K1 (wild-type and mutant) in presence of ATP (10 μM), and re-purified using Ni-NTA purification kit (Thermo-Scientific). Purified NSAP1, phospho-L13a, and GAPDH were generated as described (Jia et al., 2008 (link)). The GAIT complex was reconstituted by incubating 5 pmol each of purified EPRS linker, NSAP1, GAPDH, and phospho-L13a. The reconstituted complex was incubated with Luc-Cp-GAIT (200 ng) and T7 gene 10 (200 ng) template transcripts, and added to wheat germ extract system (Promega) containing Met-free amino acid mixture and 35S-Met (Perkin-Elmer). Translation of Luc-Cp-GAIT and T7 gene 10 transcripts was visualized by resolution on 10% SDS-PAGE and autoradiography.
Arif A., Jia J., Willard B., Li X, & Fox P.L. (2019). Multisite phosphorylation of S6K1 directs a kinase phospho-code that determines substrate selection. Molecular cell, 73(3), 446-457.e6.
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