Product Class: Other

Poly(U) Polymerase
NEB2 recombinant 37 65 Heat

Product Introduction

Poly(U) Polymerase catalyzes the template independent addition of UMP from UTP or AMP from ATP to the 3´ end of RNA.

  • Isolated from a recombinant source
  • Supplied with 10X reaction buffer
  • Tested for the absence of DNases and RNases
Catalog # Size Concentration
M0337S 60.0 units 2000 units/ml

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Product Information

Description

Poly(U) Polymerase catalyzes the template independent addition of UMP from UTP or AMP from ATP to the 3´ end of RNA.

Product Source

An E. coli strain that carries the cloned poly(U) polymerase gene of Schizosaccharomyces pombe Cid1.
This product is related to the following categories:
cDNA Synthesis & Reverse Transcriptases Products,
RNA Synthesis In vitro Transcription (IVT),
This product can be used in the following applications:
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that incorporates 1 nmol of UMP into RNA in a 50 µl volume in 10 minutes at 37°C.

Reaction Conditions

1X NEBuffer™ 2
Supplement with 0.5 mM UTP 
Incubate at 37°C

1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

65°C for 20 minutes

Unit Assay Conditions

1X NEBuffer 2, 0.5 mM 3H UTP and 5 µg yeast RNA are combined in a 50 µl reaction incubated at 37°C for 10 minutes.

Application Features

  • Labeling of RNA with UTP
  • Poly(U) and Poly(A) tailing of RNA for cloning
  • Studying effects of poly(U) tailing on stability and translation of RNA transferred into eukaryotic cells
  • Poly (A) tailing of 2' O-Me modified 3' ends

Product Notes

  1. UTP is not included with this product.
  2. Poly(U) Polymerase in NEBuffer 2 will incorporate NMP from NTP into RNA. Tailing length of poly(U) varies with UTP. Poly(U) Polymerase is highly processive under low primer concentrations (<100 pmol).

References

  1. Wickens, M. and Kwak, J.E. (2008). Science. 319, 1344.
  2. Kwak, J.E. and Wickens, M. (2008). RNA. 13, 860.
  3. Rissland, O.O.S., Mikulasova, A. and Norbury, C.J. (2007). Molecular and Cell Biology. 27, 3612.

Protocols, Manuals & Usage

Protocols

  1. A Typical Tailing Reaction (M0337)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the molecular weight of Poly(U) Polymerase?
  2. What type of labeled-UTPs will the Poly(U) Polymerase incorporate?
  3. Will Poly(U) Polymerase incorporate GTPs onto the 3’ end of an RNA?
  4. What Poly(U) length can be expected to be incorporated by Poly(U) Polymerase?
  5. Can Poly(U) Polymerase incorporate GTPs onto an RNA that contains chemically modified or 2-O-methyl groups on its 3’ end?
  6. How can the Poly(U) Polymerase be inactivated without heating up the reaction?
  7. Does the E. coli Poly(U) Polymerase work in the M-MuLV reverse transcriptase buffer?
  8. Can Poly(U) Polymerase be used to add a poly U tail to ssDNA?

Tech Tips

We strongly recommend wearing gloves, using nuclease-free tubes and reagents, and thoroughly cleaning pipettes and bench surfaces to avoid RNase contamination.

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

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

Legal and Disclaimers

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.

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