HiScribe® SP6 RNA Synthesis Kit

Catalog #	Concentration	Size	List Price	Quantity	Your Price
E2070S		50 reactions	$445.00		$400.50

Catalog #	Size	List Price	Your Price
E2070S	50 reactions	$445.00	$400.50

The HiScribe SP6 RNA Synthesis Kit is designed for the in vitro transcription of RNA using SP6 RNA Polymerase. This kit is suitable for synthesis of high yield RNA transcripts and for incorporation of cap analogs (not included) or modified nucleotides (not included) to obtain capped, biotin-labeled or dye-labeled RNA. The kit is also capable of synthesizing high specific activity radiolabeled RNA for use as probes or targets.

RNA synthesized from this kit is suitable for many applications including RNA structure and function studies, ribozyme biochemistry, probes for RNase protection or gel shift assays, hybridization-based blots, anti-sense RNA or RNAi experiments, microarray analysis, microinjection, sgRNA synthesis and in vitro translation studies.

This kit contains sufficient reagents for 50 reactions of 25 µl each. Each standard reaction yields ≥ 80 µg of RNA from 1 µg SP6 Control Template DNA. Each kit can yield ≥ 4 mg of RNA.

Figure 1 Transcription by SP6 RNA Polymerase

NEB HiScribe SP6 RNA Synthesis Kit: Comparison of RNA yields — Reactions with 1 μg linearized plasmid DNA templates were incubated at 37°C in a PCR machine according to the manufacturer’s recommendations. Transcripts were treated with DNase I, purified by LiCl precipitation and quantified using a NanoDrop™ Spectrophotometer. The HiScribe SP6 RNA Synthesis Kit from NEB is faster and achieves higher yields for RNAs of various lengths.

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

E2070S -20

SP6 Reaction Buffer	B2070AVIAL	-20	1 x 0.125 ml
SP6 RNA Polymerase Mix	M2070AVIAL	-20	1 x 0.125 ml
SP6 Control Template	N2070AVIAL	-20	1 x 0.02 ml	0.25 mg/ml
ATP (Tris)	N2071AVIAL	-20	1 x 0.125 ml	50 mM
GTP (Tris)	N2072AVIAL	-20	1 x 0.125 ml	50 mM
UTP (Tris)	N2073AVIAL	-20	1 x 0.125 ml	50 mM
CTP (Tris)	N2074AVIAL	-20	1 x 0.125 ml	50 mM
DNase I (RNase-free)	M0303AVIAL	-20	1 x 0.1 ml	2,000 units/ml
LiCl Solution	B2051AVIAL	-20	1 x 1.4 ml
Dithiothreitol (DTT)	B1222AVIAL	-20	1 x 0.5 ml	100 mM

Properties & Usage

Materials Required but not Supplied

DNA Template: The DNA template must be linear and contain the SP6 RNA Polymerase promoter with the correct orientation in relation to the target sequence to be transcribed.
General: Thermocycler or 37°C dry air incubator, microcentrifuge, nuclease-free water, nuclease-free tubes and tips
Purification: Phenol, chloroform, ethanol, 3 M Sodium Acetate, pH 5.2 or 5 M Ammonium Acetate, spin columns, equipment for RNA quantitation
Gel Analysis: Gels, running buffers, loading dye, nucleic acid ladders, gel apparatus, power supply

Optional Materials
Cap Analogs: NEB #S1411, #S1404, #S1405, #S1406, #S1407
Modified NTPs: Biotin-, Fluorescein-, Digoxigenin-, Aminoallyl-
Labeling: [α-³²P] labeled ribonucleotide (800-6,000 Ci/mmol), storage phosphor screen

Companion Products

Protocols

DataCards

manualE2070

FAQs

Citations

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

Legal And Disclaimer

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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.