Product Class: Other

Bsu DNA Polymerase, Large Fragment
NEB2 cloned at NEB recombinant 37 75 Heat

Product Introduction

Optimal performance in a number of applications

  • Strand displacement DNA synthesis
  • Random primer labeling
  • Second strand cDNA synthesis

Catalog # Size Concentration
M0330S 200.0 units 5000 units/ml
M0330L 1000.0 units 5000 units/ml

Product Information

Description

Bsu DNA Polymerase I, Large Fragment retains the 5´→ 3´ polymerase activity of the Bacillus subtilis DNA polymerase I (1), but lacks the 5´→ 3´ exonuclease domain. This large fragment naturally lacks 3´→ 5´ exonuclease activity.


Product Source

An E. coli strain that contains a genetic fusion of the Bacillus subtilis DNA polymerase I gene (starting from codon 297 thus lacking the 5´→ 3´ exonuclease domain), and the gene coding for maltose binding protein (MBP). The fusion protein is purified to near homogeneity and the MBP portion is cleaved off in vitro. The remaining DNA polymerase is purified free of MBP.
This product is related to the following categories:
DNA Manipulation Products,
This product can be used in the following applications:
Recombinase Polymerase Amplification and SIBA,
Polymerases for DNA Manipulation,
RT-PCR & cDNA Synthesis,
Isothermal Amplification,
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 will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.

Reaction Conditions

1X NEBuffer™ 2
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)

Heat Inactivation

75°C for 20 minutes

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Strand Displacement

+

Unit Assay Conditions

1X NEBuffer 2, 33 µM dNTPs including [3H]-dTTP and 70 µg/ml denatured herring sperm DNA.

Application Features

  • Random primer labeling
  • Second strand cDNA synthesis
  • Single dA tailing
  • Strand displacement DNA synthesis (2)

Product Notes

  1. Bsu DNA Polymerase, Large Fragment is not suitable for generating blunt ends because it lacks the 3´→ 5´ exonuclease necessary to remove non-templated 3´ additions.
  2. Bsu DNA Polymerase, Large Fragment retains 50% activity at 25°C and is twice as active as Klenow Fragment (3´→ 5´ exo–) at this temperature.

References

  1. Okazaki, T. et al. (1964). J. Biol.Chem.. 239, 259-268.
  2. Piepenburg, O. et al. (2006). PLOSBiology. 4, 1115-1121.

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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 [email protected].

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.