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BclI-HF

We are excited to announce that we are in the process of switching all reaction buffers to be BSA-free. Beginning April 2021, we will be gradually transitioning to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. All information on the website has been updated to reflect this change. Find more details at www.neb.com/BSA-free.

NEB high fidelity restriction endonuclease that recognizes the sequence T^GATCA

Isoschizomers | Single Letter Code | Pronunciation:

High-Fidelity (HF^®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart^™ Buffer). All HF-restriction enzymes come with Gel Loading Dye, Purple (6X). Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:

Engineered for improved performance
100% activity in rCutSmart Buffer
Time-Saver™ qualified for digestion in 5-15 minutes
Reduced star activity
Supplied with 1 vial of Gel Loading Dye, Purple (6X)
Cut Site: T/GATCA

Catalog #	Concentration	Size
R3160S	20,000 units/ml	3,000 units
R3160L	20,000 units/ml	15,000 units
Please enter a quantity for at least one size

Product Information

High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

Product Source

An E. coli strain that carries the cloned and modified BclI gene from Bacillus caldolyticus (A. Atkinson).

Reagents Supplied

The following reagents are supplied with this product:

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

R3160S -20

BclI-HF R3160SVIAL -20 1 x 0.15 ml 20,000 units/ml

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

Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

R3160L -20

BclI-HF R3160LVIAL -20 1 x 0.75 ml 20,000 units/ml

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

Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

Product Categories:: High-Fidelity (HF®) Restriction Endonucleases Products,; Restriction Endonucleases B,; time-saver-qualified-restriction-enzymes Products

Applications:: Fast Cloning: Accelerate your cloning workflows with reagents from NEB,; Restriction Enzyme Digestion

Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam^–) 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: 100%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent B
Storage Buffer

10 mM Tris-HCl
300 mM NaCl
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: Blocked
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Isoschizomers

BclI
FbaI
Ksp22I
Related Products
Companion Products
Materials Sold Separately
- rCutSmart™ Buffer
Product Notes
1. BclI-HF has the same specificity as BclI (NEB #R0160), but it has been engineered for reduced star activity.
2. Cleaves to leave a 5´ GATC extension which can be efficiently ligated to DNA fragments generated by BamHI, BglII, MboI, Sau3AI and BstYI.
3. This enzyme is blocked by dam methylation. More information can be found at Dam-Dcm and CpG Methylation.