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MboII
Available in FastDigest® format

5'-G A A G A(N)8^-3' 3'-C T T C T(N)7^-5'		digested lambda DNA (dam-) 1.0% agarose
#ER0821 Supplied with: 10X Buffer B 10X Buffer Tango™	300 u   1 ml 1 ml
#ER0822 Supplied with: 10X Buffer B 10X Buffer Tango™	1500 u   1 ml 1 ml
Commercial Isoschizomers: -
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER0821, #ER0822 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, %
				B  (blue)	G  (green)	O  (orange)	R  (red)	Tango™  (yellow)
				1X	1X	1X	1X	1X	2X
Buffer B	37°C	1.0	65°C	100	50-100	20-50	0-20	50-100	20-50

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

Ligation and Recleavage
After 5-fold overdigestion with MboII, more than 80% of the digested DNA fragments can be ligated in the reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. More than 80% of these can be recut.

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

Note

Assayed using lambda DNA (dam-) (#SD0021). Greater than 15-fold overdigestion with MboII may result in star activity. MboII may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid an atypical DNA band pattern, use the 6X Loading Dye & SDS Solution (#R1151) for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal buffer	Enzyme activity, %
	Tango™  (yellow)
	1X	2X
Buffer B	50-100	20-50

Number of Recognition Sites in DNA Molecules

Lambda	PhiX174	M13mp18/19	pBR322	pUC18/19	pUC57	pTZ19R/U	pBluescriptIIKS(-/+)	pBluescriptIISK(-/+)	pACYC177	pACYC184
130	11	11	11	7/8	8	9	8	8	15	11

Updated balandžio 22, 2008 11:29