Bsu DNA Polymerase, Large Fragment

Product Introduction

Optimal performance in a number of applications

Recombinase polymerase amplification
Strand displacement DNA synthesis
Random primer labeling
Second strand cDNA synthesis

Bioz Badge Exists : True

Catalog #	Size	Concentration
M0330S	200 units	5000 units/ml
M0330L	1000 units	5000 units/ml

Product Information

Description

Bsu DNA Polymerase, Large Fragment retains the 5´→ 3´ polymerase activity of the Bacillus subtilis DNA polymerase (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 the Bacillus subtilis DNA polymerase gene (starting from codon 297 thus lacking the 5´→ 3´ exonuclease domain).

This product is related to the following categories:: DNA Manipulation,

This product can be used in the following applications:: Recombinase Polymerase Amplification and SIBA,; Polymerases for DNA Manipulation,; RT-PCR & cDNA Synthesis,; Isothermal Amplification,

Reagents Supplied

The following reagents are supplied with this product:

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

NEB #			Stored at (°C)
M0330S			-20

	Component Name	Component #	Stored at (°C)	Amount	Concentration
	Bsu DNA Polymerase, Large Fragment	M0330SVIAL	-20	1 x 0.04 ml	5,000 units/ml
	NEBuffer™ 2	B7002SVIAL	-20	1 x 1.25 ml	10 X

NEB #			Stored at (°C)
M0330L			-20

	Component Name	Component #	Stored at (°C)	Amount	Concentration
	Bsu DNA Polymerase, Large Fragment	M0330LVIAL	-20	1 x 0.2 ml	5,000 units/ml
	NEBuffer™ 2	B7002SVIAL	-20	1 x 1.25 ml	10 X

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

3' - 5' Exonuclease

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

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.
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

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

Tools & Resources

Selection Charts

DNA Polymerase Selection Chart

Citations & Technical Literature

Citations

Additional Citations

Hoser MJ, Mansukoski HK, Morrical SW, Eboigbodin KE (2014) Strand Invasion Based Amplification (SIBA®): A Novel Isothermal DNA Amplification Technology Demonstrating High Specificity and Sensitivity for a Single Molecule of Target Analyte PLoS One; 9(11), e112656. PubMedID: 25419812, DOI: 10.1371/journal.pone.0112656

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]

M0330S_L_v2

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.

<em>Bsu</em> DNA Polymerase, Large Fragment

NEBuffer™ 2

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.

Bsu DNA Polymerase, Large Fragment

Product Introduction