SP6 RNA Polymerase

Reagents Supplied

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0207S     -20       SP6 RNA Polymerase M0207SVIAL -20 1 x 0.1 ml 20,000 units/ml   RNAPol Reaction Buffer B9012SVIAL -20 1 x 1 ml 10 X

Application Features

• Radiolabeled RNA probe preparation
• Non-isotopic RNA labeling
• Preparation for RNA vaccines
• Guide RNA for gene targeting
• mRNA for in vitro translation and micro injection
• RNA structure, processing and catalysis studies
• RNA amplification
• Anti-sense RNA for gene expression experiment

Companion Products

• dsRNA Ladder
• Ribonucleotide Solution Mix
• ssRNA Ladder
• T7 RNA Polymerase
• Pyrophosphatase, Inorganic (E. coli)
• RNA Loading Dye, (2X)
• E. coli Poly(A) Polymerase
• RNase Inhibitor, Murine
• Thermostable Inorganic Pyrophosphatase
• mRNA Cap 2 O Methyltransferase
• Vaccinia Capping System

1. For radio labeled high specific activity RNA probes, the concentration of the radioactive nucleotide should be limited to 6 μM. 
2. To protect RNA against ribonuclease, RNase inhibitor (NEB #M0314 or #M0307) should be added to a final concentration of 1 U/μl. 
3. SP6 RNA Polymerase is extremely sensitive to salt inhibition. The overall salt concentration should not exceed 50 mM. 
4. SP6 RNA Polymerase is slightly more active at 40°C than at 37°C. Incubation at 40°C may be considered for RNA transcript containing strong secondary structures. 

1. Schenborn, E.T. and Meirendorf, R.C. (1985). Nucl. Acids Res. 13, 6223-6236.
2. Zinn, K. et al. (1983). Cell. 34, 865-879.
3. Butler, E.T. and Chamberlin, J. (1982). J. Biol. Chem. 257, 5772-5778.
4. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual . (2nd ed.), 10.27-10.37.
5. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 18.82-18.84.
6. Melton, D.A., Krieg, P.A., Rebagliati, M.R., Maniatis, T., Zinn, K. and Green, M.R. (1984). Nucl. Acids Res. 12, 7057-7070.
7. Kreig, P.A. and Melton, D.A. (1984). Nucl. Acids Res. 12, 7057-7070.
8. Green, M.R., Maniatis, R. and Melton, D.A. (1983). Cell. 32, 681-694.
9. Melton, D.A. (1985). Proc. Natl. Acad. Sci. USA. 82, 144-128.
10. Milligan, J.F., Groebe, D.R., Witherell, G.W. and Uhlenbeck, O.C. (1987). Nucl. Acids Res. 15, 8783.

1. SP6 In Vitro Transcription Reaction Protocol (M0207)

• DNA Polymerase Selection Chart
• RNA Polymerase Selection Chart

1. Does the transcription reaction with SP6 RNA Polymerase require a primer?
2. Does SP6 RNA Polymerase leave an extra base at the end of a transcript?
3. Will SP6 RNA Polymerase work on single stranded substrate?
4. Will SP6 RNA Polymerase work on uncut plasmid DNA?
5. Can aberrant RNA be produced when using SP6 RNA Polymerase?
6. Can I use SP6 RNA Polymerase to make high specific activity radiolabeled probes?
7. Why is the specific activity of the probe low?
8. What are the main causes of reaction failure using SP6 RNA Polymerase?

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 & Change Notifications

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]

• M0207S_L_v1

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]

• M0207S_L_v1_0151802
• M0207S_v1_10018458
• M0207L_v1_10020002
• M0207S_v1_10034633
• M0207S_v1_10039728
• M0207L_v1_10039730
• M0207S_v1_10052055
• M0207S_v1_10058293
• M0207S_v1_10062864
• M0207S_v1_10077617
• M0207S_v1_10086517
• M0207S_v1_10094846
• M0207S_v1_10110024
• M0207S_v1_10131482
• M0207S_v1_10164670

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• SP6 RNA Polymerase
• RNAPol Reaction Buffer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.