HiScribe™ T7 High Yield RNA Synthesis Kit
- Up to 180 μg of RNA per reaction from 1 μg of control template
- Enables full substitution of NTPs for labeling and incorporation of modified bases
- Linearized control template included for verification of RNA synthesis
Avoiding RNase Contamination
The HiScribe T7 High Yield RNA Synthesis Kit is an extremely flexible system for in vitro transcription of RNA using T7 RNA Polymerase. The kit allows for synthesis many kinds of RNA including internally labeled and co-transcriptionally capped transcripts.
RNA synthesized from the kit is suitable for many applications including RNA structure and function studies, ribozyme biochemistry, probes for RNase protection assays and hybridization based blots, anti-sense RNA and RNAi experiments, microarray analysis, microinjection, and in vitro translation and RNA vaccines.
The kit contains sufficient reagents for 50 reactions of 20 μl each. Each standard reaction yields up to 180 μg of RNA from 1 μg control template. Each kit can yield up to 9 mg RNA. For 32P labeling, the kit contains enough reagents for 100 reactions of 20 μl each.
Materials Not Included:
- DNA Template: The DNA template must be linear and contain the T7 RNA Polymerase promoter with correct orientation in relation to target sequence to be transcribed.
- 3′-O-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (NEB #S1411)
- m7G(5′)ppp(5′)G RNA Cap Structure Analog (NEB #S1404)
- m7G(5′)ppp(5′)A RNA Cap Structure Analog (NEB #S1405)
- G(5′)ppp(5′)A RNA Cap Structure Analog (NEB #S1406)
- G(5′)ppp(5′)G RNA Cap Structure Analog (NEB #S1407)
- Modified-NTP: Biotin-, Fluorescein-, Digoxigenin-, or Aminoallyl-NTP
- Labeling: [α-32P] labeled ribonucleotide (800-6,000 Ci/mmol)
- General: 37°C incubator or PCR machine, nuclease-free water
- DNase I: DNase I (RNase-free) (NEB #M0303)
- Purification: Buffer- or water-saturated phenol/chloroform, ethanol and 3 M sodium acetate, pH 5.2, spin columns
- Gel Analysis: Gels and running buffers, gel apparatus, power supply
The following reagents are supplied with this product:
Store at (°C) Concentration 10X T7 Reaction Buffer -20 10 X T7 RNA Polymerase Mix -20 FLuc Control Template -20 0.5 μg/μl ATP -20 100 mM GTP -20 100 mM UTP -20 100 mM CTP -20 100 mM
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