BcgI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10271508. Learn more.

SAM now included in enzyme formulation – no longer supplied as a separate vial.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB restriction endonuclease that recognizes the sequence _NN^NNNNNNNNNNCGANNNNNNTGCNNNNNNNNNN_NN^
Isoschizomers | Single Letter Code

Product Introduction

  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence

  • Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.

  • Restriction Enzyme Cut Site: (10/12)CGANNNNNNTGC(12/10)

Featured Videos

View Video Library
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    Reduce Star Activity with High-Fidelity Restriction Enzymes

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    Standard Protocol for Restriction Enzyme Digests

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    NEB® TV Ep. 15 – Applications of Restriction Enzymes

  • cuttingclose_video_thumb

    Restriction Enzyme Digest Protocol: Cutting Close to DNA End

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    Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel

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    Why is My Restriction Enzyme Not Cutting DNA?

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    Restriction Enzyme Digest Problem: Too Many DNA Bands

  • nebcloner_dd_demo_thumb

    Double Digestion with NEBcloner

  • Type IIS Restriction Enzymes function

    Function and Application of Type IIS Restriction Enzymes

Product Information

Description

Product Source

An E. coli strain that carries the BcgI gene from Bacillus coagulans (H. Kong).
This product is related to the following categories:
Restriction Endonucleases B
This product can be used in the following applications:
Restriction Enzyme Digestion

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 λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ r3.1
Incubate at 37°C

1X NEBuffer™ r3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

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

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml Recombinant Albumin
0.32 mM S-adenosylmethionine (SAM)
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

dam methylation: Impaired by Overlapping
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Some Combinations of Overlapping

Product Notes

  1. BcgI cleaves DNA substrates twice to excise its recognition site generating a 32 base-pair fragment with 2-base 3´ overhangs.
  2. May exhibit Star Activity in NEBuffer r2.1 and rCutSmart Buffer.
  3. Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.
  4. Impaired by overlapping dam methylation and blocked by some combinations of overlapping CpG methylation.
  5. BcgI requires S-adenosylmethionine (SAM) for optimal activity

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. Does BcgI require SAM for activity?
  2. Do BcgI generated ends ligate?
  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.

Specification Change Notifications

BcgI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10271508. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

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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