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Esp3I (BsmBI)
 Available in FastDigest® format

5'-C G T C T C(N)1^-3'
3'-G C A G A G(N)5^-5'

digested lambda DNA
1.0% agarose
         
#ER0451
 Supplied with:
10X Buffer Tango™		 200 u
 
1 ml
#ER0452
 Supplied with:
10X Buffer Tango™		 1000 u
 
1 ml
Commercial Isoschizomers: BsmBI

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER0451, #ER0452
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 Tango™ + 
1.0 mM DTT 		37°C 0.2 65°C 100
(+DTT)		 20-50
(+DTT)		 0-20
(+DTT)		 0-20
(+DTT)		 100
(+DTT)		 0-20
(+DTT)

Storage Buffer
10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM KCl, 1 mM EDTA, 7 mM 2-mercaptoethanol, 0.5 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 50-fold overdigestion with Esp3I, 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.

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 enzyme requires DTT (#R0861/2). Freshly made DTT should be added to the reaction buffer.
  • Esp3I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for PCR product cloning.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer Tango™ + 
1.0 mM DTT 		100
(+DTT)		 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
14 0 1 1 2 2 0 0 0 1 2

See also General Properties of Restriction Enzymes:

 

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Updated balandžio 22, 2008 11:19