CspCI

SAM now included in enzyme formulation – no longer supplied as a separate vial.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB restriction endonuclease that recognizes the sequence _NN^NNNNNNNNNNNCAANNNNNGTGGNNNNNNNNNN_NN^

Isoschizomers | Single Letter Code | Pronunciation:

Time-Saver™ qualified for digestion in 5-15 minutes
100% activity in rCutSmart^™ 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.
Restriction Enzyme Cut Site: (11/13)CAANNNNNGTGG(12/10)

Catalog #	Concentration	Size	List Price	Your Price
R0645S	5,000 units/ml	500 units	$74.00	Sign In Sign In

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Product Information

Product Source

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

This product is related to the following categories:: Restriction Endonucleases C G,; Time-Saver Qualified Restriction Enzymes Products

This product can be used in the following applications:: Restriction Enzyme Digestion

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

R0645S -20

CspCI R0645SVIAL -20 1 x 0.1 ml 5,000 units/ml

rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

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 rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Usage Concentration
5,000 units/ml

Activity in NEBuffers
NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent A
Storage Buffer

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

Heat Inactivation
65°C for 20 minutes
Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Related Products
Companion Products
Materials Sold Separately
- rCutSmart™ Buffer
Product 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. 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.
4. Not sensitive to CpG, dcm, or dam methylation.
5. CspCI requires S-adenosylmethionine (SAM) in reaction mixture for optimal activity (SAM is supplied in the enzyme formulation).