HphI

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0158S     -20       HphI R0158SVIAL -20 1 x 0.2 ml 5,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

Companion Products

• dam^-/dcm^- Competent E. coli

Materials Sold Separately

• rCutSmart™ Buffer

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%.

1. Optimizing Restriction Endonuclease Reactions
2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
3. Restriction Digest Protocol
4. 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
• Dam and Dcm Methylases of E. coli
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Effects of CpG Methylation on Restriction Enzyme Cleavage
• Heat Inactivation
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Restriction Enzymes for Droplet Digital PCR (ddPCR)
• Single Letter Codes
• Star Activity

• 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
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Type IIS Restriction Enzymes

• DNA Sequences and Maps Tool

• Enzyme Finder
• NEBioCalculator®
• REBASE®

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?

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.

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]

• R0158S_L_v3

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]

• R0158S_L_v1_1001211
• R0158S_L_v1_1011305
• R0158S_L_v1_1011311
• R0158S_L_v1_1011405
• R0158S_L_v1_1011410
• R0158S_L_v1_1011504
• R0158S_L_v1_1011509
• R0158S_L_v1_1011510
• R0158S_L_v1_1011602
• R0158S_L_v1_1011603
• R0158S_L_v1_1011609
• R0158S_L_v2_1011707
• R0158S_L_v2_1011802
• R0158L_v2_10010513
• R0158S_v2_10010515
• R0158L_v2_10030514
• R0158S_v2_10030516
• R0158L_v2_10041270
• R0158S_v2_10049613
• R0158L_v2_10055333
• R0158S_v2_10063033
• R0158L_v2_10073970
• R0158S_v2_10076439
• R0158L_v2_10089309
• R0158S_v2_10089307
• R0158S_v2_10091707
• R0158L_v2_10100721
• R0158S_v2_10100720
• R0158L_v2_10118758
• R0158L_v3_10128093
• R0158S_v3_10128092
• R0158S_v3_10157698
• R0158L_v3_10164031
• R0158L_v3_10179989
• R0158S_v3_10179991
• R0158S_v3_10212851
• R0158L_v3_10226027
• R0158S_v3_10242812
• R0158L_v3_10250726
• R0158S_v3_10276641
• R0158L_v3_10277422
• R0158S_v3_10289650
• R0158L_v3_10289667
• R0158S_v3_10324878

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

• HphI
• rCutSmart™ Buffer

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.