Product Class: Restriction Endonuclease

HphI
neb31 cloned at NEB recombinant time saver dil_B 37 65 Heat dam dcm

HphI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10128083. Learn more

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 GGTGANNNNNNN_N^
Isoschizomers | Single Letter Code

Product Introduction

  • Time-Saver™ qualified for digestion in 5-15 minutes
  • 100% activity in rCutSmart™ Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
  • Restriction Enzyme Cut Site: GGTGA(8/7)
Catalog # Size Concentration
R0158S 1000.0 units 5000 units/ml
R0158L 5000.0 units 5000 units/ml

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Product Information

Description

Product Source

An E. coli strain that carries the HphI gene from Haemophilus parahaemolyticus (ATCC 49700).
This product is related to the following categories:
Restriction Endonucleases H M Products,
Time-Saver Qualified Restriction Enzymes Products,
This product can be used in the following applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
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 rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

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

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

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

Isoschizomers

AsuHPI

Product Notes

  1. It has been suggested that HphI may cleave at N9/N8 depending on the sequence between the recognition and cleavage sites (Kang, C. and Wu, C.-W. (1987) Nucl. Acids Res. 15, 2279-2294; Cho, S.-H. and Kang, C. (1990) Mol. Cells 1, 81?86.). Incubation of > 12 units for over 4 hours on ΦX174 DNA results in additional cleavage products. This has not yet been shown to occur on other DNAs. Low pH and high glycerol concentration enhance this activity.
  2. This enzyme is blocked by dcm methylation. More information can be found at Dam-Dcm and CpG Methylation.
  3. Star activity may result from extended digestion, high enzyme concentration or a glycerol concentration of >5%
  4. 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. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  3. Restriction Digest Protocol
  4. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  5. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is there variability in the cleavage site of HphI?
  2. Does HphI exhibit star activity?
  3. What is the activity of HphI  at 25°C?
  4. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  5. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
  6. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  7. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Tech Tips

This enzyme will not cut plasmids isolated from most E.coli because it is blocked by dam methylation and dcm methylation. For enzyme cleavage to occur, plasmids must be transformed and isolated from a dam-, dcm- strain of E.coli, such as NEB’s dam-/dcm- Competent E.coli (NEB #C2925).

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

HphI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10128083. All subsequent (higher number) lots will contain rAlbumin. 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 [email protected].

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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