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Eco57MI*

* This product and process are covered by
U.S. patent No. 6893854 and corresponding counterparts.


digested lambda DNA
1.0% agarose 
5'-C T G Pu A G(N)16^-3'
3'-G A C Py T C(N)14^-5'

       
#ER1671
Supplied with:
10X Buffer B
50X SAM
10X Buffer Tango™		 50 u
 
1 ml
0.1 ml
1 ml
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1671
MSDS (English)
MSDS (English-USA)
MSDS (German)
Chart (in pdf, 721 KB)
Brochure (in pdf, 316 KB)

Concentration
2 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 +
1 µM S-adenosylmethionine		 37°C 1.0 65°C 100
(+SAM)		 50-100
(+SAM)		 0-20
(+SAM)		 20-50
(+SAM)		 50-100
(+SAM)		 0-20
(+SAM)

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

Ligation and Recutting
After 10-fold overdigestion with Eco57MI, approximately 90% of the digested DNA fragments can be ligated but none of these can be recut due to the methylation of the recognition sequence by restriction enzyme.

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

Note
  • Eco57MI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine.
    1 µM S-adenosylmethionine gives more than a 100-fold increase in Eco57MI activity. Still, a complete cleavage of some substrates by Eco57MI is difficult to achieve.
  • Eco57MI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase.
  • Eco57MI 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 B +
0.01M S-adenosylmethionine		 50-100
(+SAM)		 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
65 3 2 6 3 3 3 3  4 2 6

See also General Properties of Restriction Enzymes:

 

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Updated vasario 06, 2008 14:11