I-SceI

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 TAGGG_ATAA^CAGGGTAAT

Isoschizomers | Single Letter Code | Pronunciation:

100% activity in rCutSmart^™ Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
This is a homing endonuclease
Tolerates some sequence degeneracy within recognition sequence
Restriction Enzyme Cut Site: TAGGGATAACAGGGTAAT(-9/-13)

Catalog #	Concentration	Size	List Price	Your Price
R0694S	5,000 units/ml	500 units	$74.00	Sign In Sign In
R0694L	5,000 units/ml	2,500 units	$304.00	Sign In Sign In

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Product Information

I-SceI is an intron-encoded endonuclease. The intron encoding I-SceI is present in the mitochondria of Saccharomyces cerevisiae.

Product Source

An E. coli strain that carries the I-SceI mitochondrial gene from Saccharomyces cerevisiae (B. Dujon).

Reagents Supplied

The following reagents are supplied with this product:

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

R0694S

-80

I-SceI	R0694SVIAL	-80	1 x 0.1 ml	5,000 units/ml
rCutSmart™ Buffer	B6004SVIAL	-20	1 x 1.25 ml	10 X
pGPS2 NotI-linearized Control Plasmid	N0420SVIAL	-20	1 x 0.01 ml	0.5 mg/ml

R0694L

-80

I-SceI	R0694LVIAL	-80	1 x 0.5 ml	5,000 units/ml
rCutSmart™ Buffer	B6004SVIAL	-20	1 x 1.25 ml	10 X
pGPS2 NotI-linearized Control Plasmid	N0420SVIAL	-20	1 x 0.01 ml	0.5 mg/ml

This product is related to the following categories:: Homing Endonucleases Products,; Restriction Endonucleases H M

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

Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to cleave 1 µg of pGPS2 NotI-Iinearized Control Plasmid 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)

Activity in NEBuffers
NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 50%
NEBuffer™ r3.1: 25%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent B
Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation
65°C for 20 min
Related Products
Companion Products
Product Notes
1. Homing endonucleases do not have stringently-defined recognition sequences in the way that restriction enzymes do. That is, single base changes do not abolish cleavage but reduce its efficiency to variable extents. The precise boundary of required bases is generally not known. The recognition sequence listed is one site that is known to be recognized and cleaved.
2. Supplied with plasmid DNA. NotI-linearized pGPS2 is supplied at 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0), 1 mM EDTA. Cleavage of this 2499 bp plasmid with I-SceI gives fragments of 1518 and 981 base pairs.
3. The enzyme retains full activity after at least 10 free-thaw cycles; the enzyme can be aliquoted and stored at –80C.