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AjuI
Available in FastDigest® format

5'-^ 7(N)G A A(N)7T T G G(N)11^-3'
3'-^12(N)C T T(N)7A A C C(N) 6^-5'

digested lambda DNA
0.7% agarose

        
#ER1951
Supplied with:
10X Buffer R
50X SAM
10X Buffer Tango™		 100 u
 
1 ml
0.1 ml
1 ml
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1951
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 + 0.01 mM SAM 37°C 0.5 65°C 0-20
(+SAM)		 50-100
(+SAM)		 20-50
(+SAM)		 100
(+SAM)		 50-100
(+SAM)		 50-100
(+SAM)

Storage Buffer
AjuI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/ml BSA and 50% (v/v) glycerol.

Ligation and Recleavage
After 50-fold overdigestion with AjuI, more than 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 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
  • Complete cleavage of some substrates with AjuI is difficult to achieve.
  • AjuI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase.
  • AjuI produces double-strand cuts on both sides of the interrupted recognition site. In certain sequence contexts, the cleavage position may be shifted by one base pair. However, the cleavage position indicated above will predominate.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer R + 0.01 mM SAM 50-100
(+SAM)		 50-100
(+SAM)

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
3 0 0 0 0 0 0 0 0 1 0

See also General Properties of Restriction Enzymes:

 

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Updated balandžio 24, 2008 09:52