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AloI

5'-^   7(N) G A A C (N)6 T C C (N)12-13^-3'
3'-^12-13(N) C T T G (N)6 A G G (N)7    ^-5' 
digested lambda DNA
0.7% agarose
        
#ER1491
Supplied with:
10X Buffer R
10X Buffer Tango™		 100 u
 
1 ml
1 ml
#ER1492
Supplied with:
10X Buffer R
10X Buffer Tango™		 500 u
 
1 ml
1 ml
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1491, #ER1492
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 30°C 0.1 65°C 0-20 0-20 0-20 100 20-50 100

Storage Buffer
 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% glycerol.

Ligation and Recleavage
After 10-fold overdigestion with AloI, approximately 70% of the digested DNA fragments can be ligated and more than 80% of these can be recut.

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

Note
  • Incubation at 37°C results in 20% activity.
  • AloI produces double-strand cuts on both sides from the interrupted recognition site. Its unique feature is a degenerate cleavage point on the 3’ side of the recognition sequence (12 or 13 nt away).
  • The presence of SAM in the reaction mixture results in incomplete cleavage with AloI.
  • Greater than 10-fold overdigestion with AloI may result in star activity.
  • AloI 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 R 20-50 100

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
7 0 1 0 0 0 1 1 1 0 0

See also General Properties of Restriction Enzymes:

 

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Updated kovo 06, 2008 13:04