< td> < td>3'

5'		C	C ↓	T	N	A	G	C		3'
		G	G	A	N	T	C	G		5'

Thermo Scientific Nt.Bpu10I nickase recognizes the CC^TNAGC site and cleaves best in R buffer at 37°C. For a table of enzymatic activities, double digestion conditions, and heat inactivation of this and other restriction enzymes, see Reaction conditions for restriction enzymes.

Thermo Scientific Conventional Restriction Endonucleases are a large collection of high quality restriction enzymes optimized to function in one buffer of a five-buffer system. In addition, a universal Tango buffer is provided to facilitate double enzyme digestion. All enzymes demonstrated 100% activity under recommended buffers and reaction conditions. To ensure stable endonuclease activity, Thermo Scientific restriction enzyme reaction buffers are added with BSA, which enhances the stability of multiple enzymes and binds possible contaminants in DNA preparations.

Features

• Superior quality—strict quality control and industry-leading production processes
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for dual degradation
• BSA premixed in reaction buffer
• Broad range of restriction endonuclease specificities

Applications
Production of single-stranded circular DNA from supercoiled double-stranded plasmids in vitro, which is subsequently used for DNA sequencing, site-specific mutagenesis, etc. Nested deletions to create vectors for ligation-independent cloning methods Preparation of covalently closed double-stranded linear DNA molecules

Remarks:Nt.Bpu10I may bind to severed DNA. This can cause DNA band drift during electrophoresis. To avoid atypical DNA banding patterns, use 6X DNA loading dye with SDS solution for sample preparation, or heat digested DNA in the presence of SDS before electrophoresis.