Product Class: Restriction Endonuclease

CviAII

icon_cloned icon_recomb icon_timesaver5 icon_CutSmart dil_C-grey 25 degree incubation temp icon_HI_65
CviAIIcutsite_1_v1_000015

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
Catalog # Size Concentration
R0640L 1,000 units 10,000 units/ml
R0640S 200 units 10,000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the CviAII gene from Chlorella virus PBCV-1 (J.L. Van Etten).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
CutSmart® Buffer -20 10 X
Product Categories:
Restriction Endonucleases: C-G,
Time-Saver™ Qualified Restriction Enzymes
Applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of pUC19 DNA in 1 hour at 25°C in a total reaction volume of 50 µl.

Reaction Conditions

1X CutSmart® Buffer
Incubate at 25°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: 50%
NEBuffer 2.1: 50%
NEBuffer 3.1: 10%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
0.15% Triton® X-100
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

Activity at Temperature

@37°C: 20%

Notes

  1. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
  2. CviAII is a neoschizomer of NlaIII.

Faqs & Tech Tips

FAQs

  1. Is CviAII activity sensitive to dam, dcm or mammalian CpG methylation?
  2. How many base pairs should be added at the end of a PCR primer after the CviAII recognition site to guarantee that CviAII will cut properly?
  3. Does CviAII have any neoschizomers?
  4. My CviAII enzyme was working fine several months ago, but now it is not working anymore. Is there a reason?
  5. I have run my DNA digested with CviAII in a gel and I cannot see the digested fragment that I was expecting. Has the enzyme degraded my substrate DNA?
  6. What is the activity of CviAII at 37°C?
  7. My restriction enzyme used to be available at a lower concentration. Why does it now come at a higher concentration of 10,000 u/ml?
  8. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  9. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  10. How can I access the old NEBuffer Activity Chart?
  11. 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?
  12. Why is my Restriction Enzyme not cutting DNA?
  13. Why do I see a DNA smear on an agarose gel after a restriction digest?
  14. Why do I see additional DNA bands on my gel after a restriction digest?
  15. 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
  3. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  4. Protocol for Digestion Prior to droplet digital PCR (ddPCR)

Other Tools & Resources

Selection Charts

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.