New England Biolabs Canada

HiScribe® T7 Quick High Yield RNA Synthesis Kit

Catalog # Concentration Size
E2050L 250 reactions
E2050S 50 reactions
Catalog # Concentration Size
E2050L 250 reactions
E2050S 50 reactions
  • Up to 180 μg of RNA generated per reaction, from 1 μg of control template
  • Master mix format reduces pipetting steps while still enabling partial substitution of NTPs for labeling and incorporation of modified bases including N1-Methyl-Pseudouridine-5’-Triphosphate (NEB #N0431), 5-Methyl-Cytidine-5’-Triphosphate (NEB #N0432), Pseudouridine-5’-Triphosphate (NEB #N0433) and 5-Methoxy-Uridine-5’-Triphosphate (NEB #N0434)
  • Template removal and RNA purification reagents included
  • Linearized control template included for verification of RNA synthesis
  • Getting ready to scale up RNA synthesis? Download our new technical note “Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production” for a generalized set of recommendations for synthesizing high yields of RNA.
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    Avoiding RNase Contamination
The HiScribe T7 Quick High Yield RNA Synthesis Kit is designed for quick set-up and production of large amounts of RNA in vitro. The reaction can be set up conveniently by combining the NTP buffer mix, T7 RNA Polymerase mix and a suitable DNA template. The kit also allows for capped RNA or dye-labeled RNA synthesis by incorporation of cap analog (ARCA, NEB #S1411) or dye-modified nucleotides. RNA synthesized with the kit can be used for RNA structure and function studies, ribozyme biochemistry, as probes for RNase protection assays and hybridization based blots, anti-sense RNA and RNAi experiments, microarray analysis and microinjection, as well as in vitro translation and RNA vaccines.

To synthesize high specific activity radioactive RNA probes or RNA with 100% substitution of one or more modified nucleotides we recommend using the T7 High Yield RNA Synthesis Kit (NEB #E2040), in which the four nucleotides are supplied separately.

DNA Template Preparation:
Linearized plasmid DNA, PCR products or synthetic DNA oligonucleotides can be used as templates for in vitro transcription with the HiScribe T7 Quick High Yield RNA Synthesis Kit, provided that they contain a double-stranded T7 promoter region upstream of the sequence to be transcribed. Figure 1 illustrates the minimal T7 promoter sequence, as well as a run-off transcript after T7 transcription.

Figure 1. Transcription by T7 RNA PolymeraseFigure 1. Transcription by T7 RNA Polymerase
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  FLuc Control Template N0426AVIAL -20 1 x 0.01 ml 0.5 µg/µl
  DNase I (RNase-free) M0303AVIAL -20 1 x 0.1 ml 2,000 units/ml
  T7 RNA Polymerase Mix M0255AVIAL -20 1 x 0.1 ml
  LiCl Solution B2051AVIAL -20 1 x 1.4 ml
  NTP Buffer Mix N2052AVIAL -20 1 x 0.5 ml
  Dithiothreitol (DTT) B1222AVIAL -20 1 x 0.5 ml 100 mM
  FLuc Control Template N0426AVIAL -20 1 x 0.01 ml 0.5 µg/µl
  DNase I (RNase-free) M0303SVIAL -20 1 x 0.5 ml 2,000 units/ml
  Dithiothreitol (DTT) B1222AVIAL -20 1 x 0.5 ml 100 mM
  T7 RNA Polymerase Mix M0255LVIAL -20 1 x 0.5 ml
  LiCl Solution B2051AAVIAL -20 1 x 6.5 ml
  NTP Buffer Mix N2052AAVIAL -20 2 x 1.25 ml

Properties & Usage

Materials Required but not Supplied

  • DNA Template
  • Thermocycler or 37°C incubator.
  • Nuclease-Free Water
  • Buffer- or water-saturated phenol:chloroform
  • Ethanol
  • 3 M Sodium Acetate, pH 5.2
  • 5 M Ammonium Acetate
  • Spin Columns (See Monarch® RNA Cleanup Kits, NEB #T2030, #T2040, #T2050)
  • Gels, running buffers and gel box


Companion Products
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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