Product Class: Restriction Endonuclease

XhoI

 

icon_cloned icon_recomb icon_timesaver5 icon_CutSmart dil_A-grey icon_incTemp37 icon_HI_65 icon_cpg icon_bluewhite
Xho-I-cutsite_1

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 worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality.

Catalog # Size Concentration
R0146L 25,000 units 20,000 units/ml
R0146M 25,000 units 100,000 units/ml
R0146S 5,000 units 20,000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the XhoI gene from Xanthomonas holcicola (ATCC 13461).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
CutSmart® Buffer -20 10 X
Product Categories:
Restriction Endonucleases: T-Z,
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 (HindIII digest) fragments 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: 75%
NEBuffer 2.1: 100%
NEBuffer 3.1: 100%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
50 mM KCl
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: Impaired

Notes

  1. This enzyme has shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1 μg plasmid DNA. For complete digestion of 1 μg of plasmid DNA please follow our recommended digestion protocol.
  2. XhoI is an isoschizomer of PaeR7I.

Faqs & Tech Tips

FAQs

  1. Why isn't XhoI cutting?
  2. Does XhoI produce commonly used compatible ends?
  3. How does XhoI differ from its isoschizomer, PaeR71?
  4. What is the molecular weight of XhoI?
  5. What is the activity of XhoI at 25°C?
  6. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  7. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  8. How can I access the old NEBuffer Activity Chart?
  9. Why do I see additional DNA bands on my gel after a restriction digest?
  10. Why do I see a DNA smear on an agarose gel after a restriction digest?
  11. Why is my Restriction Enzyme not cutting DNA?
  12. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  13. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

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.