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CseI (HgaI)

5'-G A C G C (N) 5^-3' 3'-C T G C G (N)10^-5'		digested pBR322 DNA 1.4% agarose
#ER1901 Supplied with: 10X Buffer R 10X Buffer Tango™	100 u   1 ml 1 ml
#ER1902 Supplied with: 10X Buffer R 10X Buffer Tango™	500 u   1 ml 1 ml
Commercial Isoschizomers: HgaI
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER1901, #ER1902 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 R	37°C	0.5	80°C (10 u)	NR	50-100*	50-100	100	100*	50-100

* Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).
NR: buffer is not recommended, because of high star activity.

Storage Buffer
10 mM Tris-HCl (pH 7.5 at 25°C), 100 mM KCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol.

Ligation and Recleavage
After 50-fold overdigestion with CseI, 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  may overlap	- no effect. - no effect. - blocked. - no effect. - effect not determined.

Note

• Assayed using pBR322 DNA (#SD0041).

• Low salt, high glycerol (>5%) concentration, pH>8.0 or large excess of the enzyme may result in star activity.
• CseI 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 R	*	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
102	14	7	11	4	4	4	4	4	5	9

Updated kovo 06, 2008 13:35