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Cfr42I (SacII)

5'-C C G C^G G-3' 3'-G G^C G C C-5'		digested lambda DNA 0.7% agarose
#ER0201 Supplied with: 10X Buffer B 10X Buffer Tango™	1200 u   1 ml 1 ml
#ER0202 Supplied with: 10X Buffer B 10X Buffer Tango™	5x1200 u   2x1 ml 1 ml
#ER0205 Supplied with: 10X Buffer B 10X Buffer Tango™	2000 u   1 ml 1 ml
Commercial Isoschizomers: KspI, SacII, Sfr303I, SgrBI, SstII
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER0201, #ER0202, #ER0205 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 B	37°C	0.1	65°C	100	50-100	0-20	0-20	50-100	0-20

Storage Buffer
10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 7 mM 2-mercaptoethanol, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 50-fold overdigestion with Cfr42I, more than 95% of the digested DNA fragments can be ligated and recut.

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

Digestion of Agarose-embedded DNA
Minimum 20 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.

Note

• Certain sites in lambda and phiX174 DNAs are difficult to cleave with Cfr42I, the same as with its prototype SacII.
• Cfr42I activity is affected by high salt concentration. Trace amounts of sodium chloride remaining in the substrate DNA after completion of upstream applications may inhibit enzyme activity and result in impaired DNA cleavage.

Double Digestions using Tango™ Buffer, DoubleDigest™

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

NR: buffer is not recommended, since enzyme activity is less than 20%.

Number of Recognition Sites in DNA Molecules

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

Updated kovo 06, 2008 14:32