RsrII

Description

Product Source

An E. coli strain that carries the RsrII gene from Rhodopseudomonas sphaeroides (S. Kaplan).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
CutSmart® Buffer-2010X

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
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

NEBuffer 1.1: 25%
NEBuffer 2.1: 75%
NEBuffer 3.1: 10%
CutSmart® Buffer: 100%

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: Blocked

Notes

  1. RsrII requires two copies of its recognition sequence for efficient cleavage to occur. If a DNA molecule (plasmid or linear) has only one copy of this recognition site, RsrII cannot digest it to completion. These two copies of recognition sites should ideally be on the same molecule to achieve 100% cleavage, however, two different molecules can be used with slightly less efficiency. For more information and recommendations on working with enzymes requiring multi-sites such as RsrII, please visit Restriction Enzyme Cleavage: ‘single-site’ enzymes and ‘multi-site’ enzymes

FAQs

  1. Do degenerate recognition sites need to be palindromic?
  2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  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 do I see a DNA smear on an agarose gel after a restriction digest?
  6. Why do I see additional DNA bands on my gel after a restriction digest?
  7. Why is my Restriction Enzyme not cutting DNA?
  8. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?

Tech Tips

This enzyme may lose activity over time. The presence of DTT is important to retain good activity.
If the activity is low, spike with DTT: 1 ul of 1M DTT to a 50 ul is ample.

Protocols

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

Selection Charts

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

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]

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]

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.