T7 Promoter In Vitro Transcription

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The Basics: In Vitro Transcription | Thermo Fisher ...

(6 days ago) A more recent use of in vitro transcription is in aRNA amplification reactions. For these reactions, transcription templates are generated from RNA by using an Invitrogen™ oligo(dT)-T7 promoter primer during reverse transcription. The cDNA is converted to a double-stranded transcription template by a second-strand synthesis reaction.

https://www.thermofisher.com/us/en/home/references/ambion-tech-support/probe-labeling-systems/general-articles/the-basics-in-vitro-transcription.html

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in vitro RNA synthesis(T7 RNA ... - Barrick Lab :: RNAT7

(8 days ago) Apr 09, 2020  · T7 in vitro transcription kit is intended for the synthesis of large amounts of unlabeled or low specific activity RNA for a variety of uses, including in vitro translation, antisense/microinjection studies, and isolation of RNA binding proteins.

https://barricklab.org/twiki/bin/view/Lab/RNAT7

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MEGAscript™ T7 Transcription Kit

(5 days ago) The MEGAscript® T7 Kit is an ultra-high yield in vitro transcription kit. The high yields are achieved by modifying typical transcription reaction conditions so that very high nucleotide concentrations can be effectively used. The T7 Enzyme Mix and the 10X Reaction Buffer are specifically calibrated for each lot and RNA polymerase.

https://www.thermofisher.com/order/catalog/product/AMB13345

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Maximizing transcription of nucleic acids with efficient ...

(6 days ago) Aug 14, 2020  · In vitro transcription using T7 bacteriophage polymerase is widely used in molecular biology. Here, we use 5′RACE-Seq to screen a randomized initially transcribed region of the T7 promoter …

https://www.nature.com/articles/s42003-020-01167-x

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Bacteriophage T7 late promoters: construction and in vitro ...

(7 days ago) Oct 25, 1982  · Restriction fragments, ending at the Hinf I site located at position -10 in the promoter from 14.8% of the T7 genome, were cloned into pBR322. This produced the deletion of either the left or the right part of the promoter. The in vitro transcription properties of these plasmids were determined.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC326922/

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Optimized RNA amplification using T7-RNA-polymerase based ...

(6 days ago) A total of 28 different oligo (dT)-T7 promoter primers were tested. Two of them showed a dramatically increased yield of RNA from the in vitro transcription reaction. The combination of the improved second-strand synthesis with the new T7 primer increased the RNA yield 60-fold compared to the yield of standard procedures.

https://pubmed.ncbi.nlm.nih.gov/15464965/

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Optimized RNA amplification using T7-RNA-polymerase based ...

(4 days ago) Nov 01, 2004  · In vitro transcription from primer T7MdTV3 produces only about one third of RNA compared to that from MluT7dTV2 (see Fig. 1). This might be caused by the short 5′ flanking sequence of the T7 promoter or it might result from the second-strand cDNA synthesis.

https://www.sciencedirect.com/science/article/pii/S0003269704006074

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General plasmids for producing RNA in vitro transcripts ...

(8 days ago) The development of in vitro transcription methods using phage RNA polymerases (RNAPs), in particular T7 RNAP, to produce in vitro RNA transcripts (1–4) opened up many new avenues to RNA researchers. However, the nucleic acid transcripts produced often bear highly heterogeneous 3′‐ends as both DNA and RNA polymerases have a tendency to fall off the template before they reach the end of the DNA, or to add non‐coded ‘n + 1’ nucleotide(s) to the 3′‐end of the nascent nucleic acid (1,2,5,6). Problems of h…

https://academic.oup.com/nar/article/31/15/e82/1108054

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In Vitro Transcription Vector (in situ hybridization ...

(7 days ago) Jul 14, 2021  · T7 promoter: A promoter for the RNA polymerase from T7 bacteriophage. Drives high-level transcription of the downstream sequence of interest. This promoter is in the opposite orientation to the SP6 promoter, and will generate a transcript which is the reverse-complement of that produced from the SP6 promoter using the same template.

https://en.vectorbuilder.com/resources/vector-system/pPp_T7_SP6.html

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Maximizing transcription of nucleic acids with efficient ...

(8 days ago) In vitro transcription using T7 bacteriophage polymerase is widely used in molecular biology. Here, we use 5'RACE-Seq to screen a randomized initially transcribed region of the T7 promoter for cross-talk with transcriptional activity.

https://pubmed.ncbi.nlm.nih.gov/32796901/

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HiScribe™ T7 In Vitro Transcription Kit | NEB

(5 days ago) Jun 13, 2013  · The HiScribe T7 In Vitro Transcription Kit is a system for the in vitro synthesis of large amounts of single-stranded (ss) or double-stranded (ds)RNA.

https://www.neb.com/products/e2030-hiscribe-t7-in-vitro-transcription-kit

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High salt transcription of DNA co-tethered with T7 RNA ...

(6 days ago) Jul 22, 2021  · To confirm that the Strep-tag® II peptide addition at the N-terminus of T7 RNA polymerase does not affect transcription activity, we performed in vitro transcription reactions using Strep-T7 RNA polymerase and promoter DNA (without a biotin tag) encoding a 24 base RNA (RNA-24) under high yield transcription conditions.

https://www.jbc.org/article/S0021-9258(21)00801-2/fulltext

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In Vitro Transcription Vector (for mRNA) - VectorBuilder

(8 days ago) Jun 25, 2021  · T7 RNAP has certain base requirements for efficient transcription initiation which have been already incorporated into the vector design. The first two nucleotides of the transcribed mRNA will be GG, corresponding to the 3’-end of the T7 promoter …

https://en.vectorbuilder.com/resources/vector-system/pPp_T7_mRNA.html

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Sauer:In vitro transcription with T7 RNA polymerase ...

(5 days ago) Jul 25, 2006  · Note that if you are designing a template for transcription, T7 RNAP has certain base requirements for transcription initiation. If possible, the first two nucleotides after the promoter should be GG (to be transcribed; CC in the template strand), as these are preferred by T7 RNAP.

https://openwetware.org/wiki/Sauer:In_vitro_transcription_with_T7_RNA_polymerase

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Making RNA probes with T7 transcription - OpenWetWare

(5 days ago) Jun 20, 2009  · In vitro T7 transcription of an RNA probe Designing primers for T7 RNA synthesis designing a suitable primer combining the T7 promoter sequence and a sequence specific part attaching a T7 promoter via PCR

https://openwetware.org/wiki/Making_RNA_probes_with_T7_transcription

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A Vector with a Single Promoter for In Vitro Transcription ...

(4 days ago) Feb 05, 2016  · However, gRNAs for microinjection into zygotes are obtained by in vitro transcription from a T7 bacteriophage promoter in a separate plasmid vector. Here, we describe the design and construction of a combined U6T7 hybrid promoter from which …

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0148362

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PSF-T7 - T7 PROMOTER PLASMID plasmid vector for molecular ...

(5 days ago) PSF-T7 - T7 promoter plasmid allows transcription of RNA using the T7 bacteriophage polymerase. If you are using an in vitro transcription kit the RNA produced from this vector will be uncapped and not poly-adenylated making it unsuitable for expression in mammalian cells however some kits are available that will add these mRNA features.

https://www.sigmaaldrich.com/US/en/product/sigma/ogs37

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(PDF) High salt transcription of DNA co-tethered with T7 ...

(7 days ago) Jul 01, 2021  · More than 70% of the RNA synthesized by T7 RNA polymerase during run-off transcription in vitro can be incorrect products, up to twice as long as the expected transcripts.

https://www.researchgate.net/publication/353396181_High_salt_transcription_of_DNA_co-tethered_with_T7_RNA_polymerase_to_beads_generates_increased_yields_of_highly_pure_RNA

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T7 RNA Polymerase – discovery and uses (in vitro ...

(7 days ago) Sep 22, 2020  · So, “in vitro transcription” is a way to make lots of RNA from a DNA template, often using the RNAP of a bacteriophage (bacteria-infecting virus) called T7, …

https://thebumblingbiochemist.com/365-days-of-science/t7-rna-polymerase-discovery-and-uses-in-vitro-transcription-protein-overexpression/

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(PDF) Synthesis of RNA by In Vitro Transcription

(8 days ago) Jan 01, 2011  · Three different types of DNA templates for in vitro transcription. In the upper panel, a circular plasmid with the insert of interest cloned between a T7 …

https://www.researchgate.net/publication/49652486_Synthesis_of_RNA_by_In_Vitro_Transcription

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Transcription of RNA templates by T7 RNA polymerase ...

(8 days ago) The bacteriophage T7 DNA-dependent RNA polymerase represents a useful model for studying transcription mechanisms. It is a monomeric enzyme which requires no auxiliary transcription factors for activity ( 1 ). The gene encoding T7 RNA polymerase has been cloned and the enzyme may be overexpressed in bacteria ( 2–4 ). Moreover, knowledge of its tertiary structure facilitates rational mutagenesis strategies ( 5). T7 RNA polymerase is a highly specific enzyme ( 6): it recognizes specifi…

https://academic.oup.com/nar/article/26/15/3550/2360082

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