Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents C A T A L O G

Bpu10I

5'-C C^T N A G C-3' 3'-G G A N T^C G-5'		digested lambda DNA 0.7% agarose
#ER1181 Supplied with: 10X Buffer Bpu10I 10X Buffer Tango™	200 u   1 ml 1 ml
#ER1182 Supplied with: 10X Buffer Bpu10I 10X Buffer Tango™	1000 u   1 ml 1 ml
Commercial Isoschizomers: -
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER1181, #ER1182 MSDS (English) MSDS (English-USA) MSDS (German) Chart (in pdf, 721 KB) Brochure (in pdf, 316 KB)

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, %
				Bpu10I (unique)	B  (blue)	G  (green)	O  (orange)	R  (red)	Tango™  (yellow)
				1X	1X	1X	1X	1X	1X	2X
Buffer Bpu10I	37°C	0.2	80°C	100	0-20	20-50*	50-100*	100*	50-100*	100*

* Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

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.

Ligation and Recleavage
After 50-fold overdigestion with Bpu10I, more than 90% of the digested pBR322 DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. No more than 50% of these can be recut due to asymmetric recognition sequence of Bpu10I. The remaining uncleaved ligation products may be cut by Eco81I (Bsu36I) and Bpu1102I (BlpI).

Methylation Effects
Dam: Dcm: CpG: EcoKI: EcoBI:	never overlaps  never overlaps  may overlap  never overlaps  may overlap	- no effect. - no effect. - no effect. - no effect. - no effect.

Note

A large excess of enzyme (>4u/ µg DNA) may result in incomplete DNA cleavage. Therefore, we recommend increasing the incubation time instead of using an excess of Bpu10I. Low salt, high glycerol (>5%) concentrations, pH >7.0 or large excess of the enzyme may result in star activity.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal buffer	Enzyme activity, %
	Tango™  (yellow)
	1X	2X
Buffer Bpu10I	50-100*	100*

* Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

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

Updated kovo 06, 2008 13:30