Product Class: Restriction Endonuclease

HgaI
neb31 cloned at NEB recombinant dil_A 37 65 Heat CpG

The large size of this product was discontinued on 12/31/2020. The small size will continue to be available.

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 GACGCNNNNN^NNNNN_
Isoschizomers | Single Letter Code

Product Introduction

  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
  • Restriction Enzyme Cut Site: GACGC(5/10)
Catalog # Size Concentration
R0154S 100.0 units 2000 units/ml

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Product Information

Description

Product Source

A E. coli strain that carries the HgaI gene from Haemophilus gallinarum (ATCC 14385)
This product is related to the following categories:
Restriction Endonucleases H M Products
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
  • R0154S     -20    
      HgaI R0154SVIAL -20 1 x 0.05 ml 2,000 units/ml
      NEBuffer™ r1.1 B6001SVIAL -20 1 x 1.25 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of ΦX174 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: 100%
NEBuffer™ r3.1: 25%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

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

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

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

Isoschizomers

CseI

Product Notes

  1. Of the over 3,000 known restriction endonucleases, HgaI is one of the few that produces extensions of more than 4 bases.
  2. Overdigestions with > 5 units of HgaI per µg of DNA is not recommended.
  3. HgaI loses activity in the absence of substrate and is essentially inactive after one hour at 37°C.
  4. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
  5. Blocked by CpG 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. Is HgaI activity sensitive to dam, dcm or mammalian CpG methylation?
  2. How many base pairs should be added at the end of a PCR primer after the HgaI recognition site to guarantee that HgaI will cut properly?
  3. Does HgaI have any neoschizomers?
  4. Does HgaI exhibit star activity?
  5. 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.

Trademarks

 

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