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Eco31I (BsaI)
Available in FastDigest® format

5'-G G T C T C(N)1^-3' 3'-C C A G A G(N)5^-5'		digested lambda DNA (dcm-) 0.7% agarose
#ER0291 Supplied with: 10X Buffer G 10X Buffer Tango™	1000 u   1 ml 1 ml
#ER0292 Supplied with: 10X Buffer G 10X Buffer Tango™	5000 u   2x1 ml 1 ml
Commercial Isoschizomers: BsaI, Bso31I, BspTNI
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER0291, #ER0292 MSDS (English) MSDS (English-USA) MSDS (German) Chart (in pdf, 721 KB) Brochure (in pdf, 316 KB)

Concentration
10 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 G	37°C	0.3	65°C	50-100	100	0-20	0-20	50-100	20-50

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 Eco31I, more than 95% of the digested DNA fragments can be ligated and recut.

Methylation Effects
Dam: Dcm: CpG: EcoKI: EcoBI:	never overlaps  may overlap  may overlap  never overlaps  may overlap	- no effect. - cleavage impaired. - cleavage impaired. - no effect. - effect not determined.

Digestion of Agarose-embedded DNA
Minimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.

Note

Assayed using lambda DNA (dcm-) (#SD0021), as one of the two Eco31I recognition sites in lambda DNA is difficult to cleave. Low salt, high glycerol (>5%) concentrations, pH >8.0 or large excess of the enzyme may result in star activity. Eco31I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for direct PCR product cloning.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal buffer	Enzyme activity, %
	Tango™  (yellow)
	1X	2X
Buffer G	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
2	0	0	1	1	1	1	1	1	1	0

Updated balandžio 22, 2008 11:21