Product Class: Other

RecJf
NEB2 cloned at NEB recombinant 37 65 Heat

Product Introduction

  • DNA specific exonuclease
  • Catalyzes the removal of nucleotides from linear single-stranded DNA in the 5' to 3' direction
  • Preferred substrate is double-stranded DNA with 5' single stranded overhangs > 6 nucleotides long
  • Need help finding the right exonuclease for your experiments? Try Exo Selector 

RecJf can be used for:

  • Degradation of single-stranded DNA from the 5'-end
  • Removal of 5' protruding single-stranded termini at the ends of linear double-stranded DNA (blunt ends are not exclusively created)
Catalog # Size Concentration
M0264S 1000.0 units 30000 units/ml
M0264L 5000.0 units 30000 units/ml

Product Information

Description

RecJf is a single-stranded DNA specific exonuclease that catalyzes the removal of deoxy-nucleotide monophosphates from DNA in the 5' → 3' direction (1). RecJf is a recombinant fusion protein of RecJ and maltose binding protein (MBP). It has the same enzymatic properties as wild-type RecJ. Fusion to MBP enhances RecJf solubility (2).

When DNA containing a 22 base 5' extension is used as a substrate for RecJf, the resulting products are a mixture of DNA fragments that have blunt-ends, 5' extensions (1-5 nucleotides) and recessed 5' ends (1-8 nucleotides) (3). RecJf does not require a 5' phosphate (3).

Product Source

RecJf is overexpressed and purified as a C-terminal fusion to maltose-binding protein (MBP). MBP does not affect the catalytic activity of RecJf but does enhance RecJf solubility (2).
This product is related to the following categories:

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 required to produce 0.05 nmol TCA soluble deoxyribonucleotide in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X NEBuffer 2 with 1.5 µg sonicated single-stranded [3H]-labeled E. coli DNA.

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)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Product Notes

  1. Phosphorothioate (PT) bond is resistant to RecJf  cleavage. However, only one of the two chiral forms of the PT bond is resistant. To achieve complete protection against degradation at all PT bond positions, it is recommended to include five consecutive PT bonds in chemically synthesized DNA. Learn more here.

References

  1. Lovett, S.T., Kolodner, R.D. (1989). Proc. Natl. Acad. Sci. USA. 86, 2627-2631.
  2. Lovett, S. and Whitaker, R. Unpublished observation
  3. Whitaker, R. Unpublished observation

Protocols, Manuals & Usage

Protocols

  1. Protocol for RecJf (NEB #M0264)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the activity of RecJf in the NEBuffers?
  2. Can DNA be blunted using RecJf?
  3. What is the difference between RecJf and Exonuclease I (NEB# M0293)?
  4. Can RecJf be heat inactivated?
  5. Will RecJF work in rCutSmart buffer?

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.

Other Products You May Be Interested In