5'

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C??

C?↓

T??

N??

A??

G??

C??

?

3'

?

3'

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G??

G??

A??

N??

T??

C??

G??

?

5'

?

Thermo Scientific Nt.Bpu10I nicking enzyme recognizes CC^TNAGC sites and nicks best at 37°C in R buffer. See

Reaction Conditions for Restriction Enzymes

for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

? Superior quality—stringent quality control and industry leading manufacturing process
? Convenient color-coded Five Buffer System
? Includes universal Tango buffer for double-digestions
? BSA premixed in reaction buffers
? Wide selection of restriction endonuclease specificities

Applications
Production of single-stranded circular DNA from supercoiled double-stranded plasmids in vitro with subsequent use in DNA sequencing, site-specific mutagenesis, etc Creation of nested deletions Vector preparation for ligation independent cloning method Preparations of covalently closed, double- stranded linear DNA molecules

Note: Nt.Bpu10I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.