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AlfI

5'-^10(N)G C A(N)6T G C(N)12^-3'
3'-^12(N)C G T(N)6A C G(N)10^-5'

digested lambda DNA
1.0% agarose

       
#ER1801
Supplied with:
10X Buffer R
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: #ER1801
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 R + 0.01 mM SAM 37°C 1.0 65°C 0-20
(+SAM)		 0-20
(+SAM)		 0-20
(+SAM)		 100
(+SAM)		 0-20
(+SAM)		 20-50
(+SAM)

Storage Buffer
10 mM Tris-HCl (pH 7.5 at 25°C), 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recutting
After 20-fold overdigestion with AlfI, more than 70% of the digested DNA fragments can be ligated but none of these can be recut due to the methylation of the recognition sequence by this enzyme.

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

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
A complete cleavage of some substrates with AlfI is difficult to achieve. 
AlfI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase. 

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer R + 0.01 mM SAM NR 20-50
(+SAM)

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
22 0 1 3 0 1 0 0 0 1 3

See also General Properties of Restriction Enzymes:

 

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Updated vasario 06, 2008 13:31