HgaI

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

Companion Products

• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

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%

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

• Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Effects of CpG Methylation on Restriction Enzyme Cleavage
• Heat Inactivation
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

• Alphabetized List of Recognition Sequences
• Compatible Cohesive Ends and Generation of New Restriction Sites
• Dam-Dcm and CpG Methylation
• Enzymes with Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Type IIS Restriction Enzymes
• Why Choose Recombinant Enzymes?

• Competitor Cross-Reference Tool
• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter™ v3.0
• NEBioCalculator®
• REBASE®

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. Why is my Restriction Enzyme not cutting DNA?
6. Why do I see a DNA smear on an agarose gel after a restriction digest?
7. Why do I see additional DNA bands on my gel after a restriction digest?
8. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
9. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

• Restriction Enzyme Troubleshooting Guide

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 & Change Notifications

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]

• R0154S_L_v1

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]

• R0154S_L_v1_0291301
• R0154S_L_v1_0291306
• R0154S_L_v1_0291311
• R0154S_L_v1_0291404
• R0154S_L_v1_0291410
• R0154S_L_v1_0291503
• R0154S_L_v1_0291512
• R0154S_L_v1_0291608
• R0154S_L_v1_0291703
• R0154S_L_v1_0291708
• R0154S_L_v1_0291803
• R0154S_v1_10020755
• R0154L_v1_10020757
• R0154S_v1_10034663
• R0154L_v1_10034661
• R0154S_v1_10051168
• R0154L_v1_10055615
• R0154L_v1_10061180
• R0154S_v1_10071561
• R0154S_v1_10083535
• R0154L_v1_10092648
• R0154S_v1_10091767
• R0154S_v1_10091563
• R0154S_v1_10101978
• R0154S_v1_10108463
• R0154S_v1_10126699
• R0154S_v1_10139683
• R0154S_v1_10167935

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• HgaI

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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