Product Class: Restriction Endonuclease

PmeI
icon_cloned icon_recomb icon_timesaver5 icon_CutSmart dil_A-grey icon_incTemp37 icon_HI_65 icon_cpg

Now comes supplied with 1 vial of Gel Loading Dye, Purple (6X), (B7024 )

Pme-I-cutsite_1

Product Introduction

Catalog # Size Concentration
R0560S 500 units 10,000 units/ml
R0560L 2,500 units 10,000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the PmeI gene from Pseudomonas mendocina (B. Zhou).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
Gel Loading Dye, Purple (6X) 25 6X
CutSmart® Buffer -20 10X
Product Categories:
Restriction Endonucleases P R,
time-saver-qualified-restriction-enzymes Products
Applications:
Restriction Enzyme Digestion

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 CutSmart® Buffer
Incubate at 37°C

1X CutSmart® Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml BSA
(pH 7.9 @ 25°C)

Activity in NEBuffers

CutSmart® Buffer: 100%
NEBuffer™ 1.1: 10%
NEBuffer™ 2.1: 50%
NEBuffer™ 3.1: 10%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

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

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

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

Product Notes

  1. This enzyme is shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1ug plasmid DNA. For complete digestion of 1ug plasmid DNA please follow our recommended digestion protocol.
  2. Based on the stability of the enzyme in the reaction, incubations longerthan 1 hr will not result in improved digestion, unless additionalenzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.

Protocols, Manuals & Usage

Protocols

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

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

Faqs & Troubleshooting

FAQs

  1. How can I improve PmeI digestion of genomic DNA embedded in agarose?
  2. What is Star Activity and how can it be avoided?
  3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  4. 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?
  5. Why is my Restriction Enzyme not cutting DNA?
  6. Why do I see additional DNA bands on my gel after a restriction digest?
  7. Why do I see a DNA smear on an agarose 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. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

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

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [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.