Product Class: Restriction Endonuclease

HpaI
neb31 cloned at NEB recombinant dil_A 37 No CpG

HpaI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10128074. 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 GTT_^AAC
Isoschizomers | Single Letter Code

Product Introduction

Catalog # Size Concentration
R0105S 500.0 units 5000 units/ml
R0105L 2500.0 units 5000 units/ml

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Product Information

Description

Product Source

An E. coli strain that carries the HpaI gene from Haemophilus parainfluenzae (ATCC 49669).
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

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: <10%
NEBuffer™ r2.1: 75%
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
200 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Methylation Sensitivity

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

Isoschizomers

KspAI

Product Notes

  1. May exhibit star activity in NEBuffer r2.1.
  2. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.
  3. Blocked by some combinations of overlapping CpG methylation.
  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. When is star activity a problem for HpaI?
  2. What do we know about star activity with Hpal I?
  3. How do you recomend using HpaI?
  4. Does a certain salt or salt level inhibit activity of HpaI?
  5. Does spermidine increase activity of HpaI?
  6. Is HpaI inhibited by dUTP incorporated at the site?
  7. Is HpaI active at 25°C?
  8. What is the molecular weight of HpaI?
  9. 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?
  10. What is Star Activity and how can it be avoided?
  11. How can this enzyme be inactivated?
  12. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  13. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  14. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
  15. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?

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

HpaI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10128074. 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 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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