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Nb.Bpu10I*

* This product and process are covered by U.S. patent No. 6867028 and corresponding counterparts.
5'-C C T N A G C-3' 3'-G G A N T^C G-5'
#ER1681 Supplied with: 10X Buffer R 10X Buffer Tango™	1000 u   1 ml 1 ml
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER1681 MSDS (English) MSDS (English-USA) MSDS (German) Chart (in pdf, 721 KB) Brochure (in pdf, 316 KB)

Description
Nb.Bpu10I is a site and strand specific endonuclease artificially engineered from restriction enzyme Bpu10I. It cleaves only one strand of the DNA within its recognition sequence on a double-stranded DNA substrate.

Concentration
5 u/µl

Conditions for 100% Activity

Recommended buffer	Incubation temperature	Units for  overnight  incubation, u/µg DNA	Thermal inactivation, in 20 min	Enzyme activity, %
				B  (blue)	G  (green)	O  (orange)	R  (red)	Tango™  (yellow)
				1X	1X	1X	1X	1X	2X
Buffer R	37°C	0.3	80°C	0-20	20-50	20-50	100	20-50	50-100

Storage Buffer
10 mM Tris-HCl (pH 7.5 at 25°C), 200 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.

Nicking and Cleavage

• Incubation of 10 units of enzyme with 1 µg pUC19 DNA (lacking the recognition sequence of Bpu10I) for 1 hour at 37°C in 50 µl reaction buffer results in <10% conversion to circular form.
• Incubation of 1 unit of enzyme with 1 µg pBR322 DNA for 1 hour at 37°C in 50 µl reaction buffer results in <5% conversion to linear form.

Applications

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

Number of Recognition Sites in DNA Molecules

Lambda	PhiX174	M13mp18/19	pBR322	pUC18/19	pUC57	pTZ19R/U	pBluescriptIIKS(-/+)	pBluescriptIISK(-/+)	pACYC177	pACYC184
19	7	4	1	0	0	0	0	0	5	3

 

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Updated vasario 06, 2008 15:52