Product Class: Restriction Endonuclease

KpnI
neb31 recombinant dil_A 37 No

This product has been replaced with the high fidelity version, KpnI-HF™ (NEB #R3142).

  • Catalog # R0142 was discontinued on December 15, 2021
NEB restriction endonuclease that recognizes the sequence G_GTAC^C
Isoschizomers | Single Letter Code

Product Introduction

Bioz Badge Exists : True
Catalog # Size Concentration

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Product Information

Description

KpnI has a High Fidelity version KpnI-HF® (NEB #R3142).

High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

Product Source

An E. coli strain that carries the KpnI gene from Klebsiella pneumoniae OK8 (ATCC 49790).
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 digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ r1.1
Incubate at 37°C

1X NEBuffer™ r1.1
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 100%
NEBuffer™ r2.1: 75%
NEBuffer™ r3.1: <10%
rCutSmart™ Buffer: 50%

Diluent Compatibility

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

No

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Isoschizomers

Acc65I
Asp718I
KpnI-HF

Product Notes

  1. Acc65I is an isoschizomer of KpnI.
  2. KpnI produces a 4-base 3´extension, whereas Acc65I produces a 4-base 5´ extension.
  3. May exhibit star activity in rCutSmart Buffer.
  4. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.
  5. Not sensitive to CpG, dcm, or dam methylation.
  6. star activity may result from extended digestion, high enzyme concentration or a glycerol concentration of >5%

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. How can this enzyme be inactivated?
  2. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  3. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  4. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting

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.

Licenses

Invitrogen
Licensed Under U.S. Patent No. 5,082,784

Invitrogen
Licensed Under U.S. Patent No. 5,192,675

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