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C A T A L O G
 

AarI

5'-C A C C T G C (N)4^-3'
3'-G T G G A C G (N)8^-5'

digested lambda DNA
1.0% agarose
           
#ER1581
 Supplied with:
10X Buffer AarI
10X Buffer Tango™
50X oligonucleotide		 25 u
 
1 ml
1 ml
25 µl
#ER1582
 Supplied with:
10X Buffer AarI
10X Buffer Tango™
50X oligonucleotide		 125 u
 
1 ml
1 ml
2x25 µl
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1581, #ER1582
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, %
AarI
(unique)		 B
(blue)		 G
(green)		 O
(orange)		 R 
(red)		 Tango™ 
(yellow)
1X 1X 1X 1X 1X 1X 2X
Buffer AarI
+ 0.5 µM of oligonucleotide		 37°C 1.0 65°C 100
(+oligo)		 NR
  (+oligo)		 NR
(+oligo)		 0-20
(+oligo)		 0-20
(+oligo)		 NR
(+oligo)		 50-100
(+oligo)

NR: buffer is not recommended, because of high star activity.

Storage Buffer
 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM KCl, 1 mM EDTA, 7 mM 2-mercaptoethanol, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 10-fold overdigestion with AarI, more than 95% of the digested DNA fragments can be ligated and recut.

Methylation Effects
Dam:
Dcm:
CpG:
EcoKI:
EcoBI:		  never overlaps
 never overlaps
 may overlap
 never overlaps
 may overlap		 - no effect.
- no effect.
- cleavage impaired.
- 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
  • For cleavage with AarI at least two copies of its recognition sequence are required.
  • Inclusion of 0.5 µM oligonucleotide with the AarI recognition sequence in the reaction mixture significantly improves cleavage of DNAs, especially of those with a single AarI site. Still, a complete cleavage of some substrates with AarI is difficult to achieve.
  • Greater than 10-fold overdigestion with AarI may result in star activity.
  • AarI may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band pattern, use the 6X Loading Dye & SDS Solution 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 AarI
+ 0.5 µM of oligonucleotide		 NR
(+oligo)		 50-100
(+oligo)

NR: buffer is not recommended, because of high star activity.

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
12 0 0 0 0 0 0 0 0 0 0

See also General Properties of Restriction Enzymes:

 

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Updated gruodžio 18, 2008 10:41