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TstI

5'-^ 8(N) C A C (N)6 T C C (N)12^-3' 3'-^13(N) G T G (N)6 A G G (N) 7^-5'		digested lambda DNA (dam-) 0.8% agarose
#ER1911 Supplied with: 10X Buffer R 10X Buffer Tango™	100 u   1 ml 1 ml
Commercial Isoschizomers: -
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #ER1911 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	0.1	No	0-20	0-20	0-20	100	0-20	100

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 50-fold overdigestion with TstI, more than 80% of the digested DNA fragments can be ligated and recut.

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

• The presence of SAM in a reaction mixture results in incomplete cleavage with TstI.
• Greater than 10-fold overdigestion with TstI may result in star activity.
• TstI 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.
• Assayed using lambda DNA (dam-) (#SD0021).

Double Digestions using Tango™ Buffer, DoubleDigest™

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

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
24	0	1	1	0	0	0	0	0	2	2

Updated vasario 06, 2008 16:09