Product Class: Restriction Endonuclease

CspCI

 

icon_cloned icon_recomb icon_timesaver5 icon_CutSmart 2+site dil_A-grey icon_incTemp37 icon_HI_65 sam
CspCIcutsitev2

Product Introduction

Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies.  Working continuously to be worth of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality.

  • Time-Saver™ qualified for digestion in 5-15 minutes
  • 100% activity in CutSmart® 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
  • Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.

Catalog # Size Concentration
R0645S 500 units 5,000 units/ml
R0645L 2,500 units 5,000 units/ml

Product Information

Description

Product Source

An E.coli strain that carries the CspCI gene from Citrobacter species 2144 (C. Nkenfou).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
CutSmart® Buffer -20 10 X
S-adenosylmethionine (SAM) -20 32 mM
Product Categories:
Restriction Endonucleases: C-G,
Time-Saver™ Qualified Restriction Enzymes
Applications:
Restriction Enzyme Digestion

Properties and 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
20 μM CspCI 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: 100%
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: Not Sensitive

Notes

  1. CspCI cleaves on both sides of its recognition sequence excising a ~35 base-pair fragment with 2-base 3´-overhangs.
  2. The exact positions of cleavage can vary ± one base depending on the sequence of the DNA flanking the recognition site and the digestion conditions
  3. S-adenosylmethionine or SAM is supplied as a 32 mM solution in 0.005 M sulfuric acid and 10% ethanol. Under these conditions SAM is stable for up to 9 months when stored at -20°C. 

Faqs & Tech Tips

FAQs

  1. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  2. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  3. How can I access the old NEBuffer Activity Chart?
  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 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?

Protocols & Manuals

Protocols

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

Other Tools & Resources

Selection Charts

Featured Articles

Usage Guildelines & Tips

Troubleshooting Guides

Restriction Enzyme Troubleshooting Guide

InteractiveTools

Quality & Safety

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 control's 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.