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C A T A L O G
 

BdaI

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

digested lambda DNA (dam-)
0.7% agarose

       
#ER1961
 Supplied with:
10X Buffer G
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: #ER1961
MSDS (English)
MSDS (English-USA)
MSDS (German)
Chart (in pdf, 721 KB)
Brochure (in pdf, 316 KB)

Concentration
1-3 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 G + 
0.05 mM S-adenosylmethionine		 30°C 1.0 65°C NR 100
(+SAM)		 0-20
(+SAM)		 20-50
(+SAM)		 50-100*
(+SAM)		 50-100
(+SAM)

NR: buffer is not recommended, because of high star activity.
* Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

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

Ligation and Recleavage
After 10-fold overdigestion with BdaI, 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:		  may overlap
 never overlaps
 never overlaps
 never overlaps
 never overlaps		 - blocked.
- no effect.
- no effect.
- no effect.
- no effect.

Note
  • Incubation at 37°C results in 30% activity.
  • Requires S-adenosylmethionine for activity. A complete cleavage of some substrates with BdaI is difficult to achieve.
  • BdaI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase.
  • BdaI 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.
  • Greater than 40-fold overdigestion with BdaI may result in star activity.
  • BdaI 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.
  • Assayed using lambda DNA (dam-) (#SD0021).

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer G + 
0.05 mM S-adenosylmethionine		 50-100*
(+SAM)		 50-100
(+SAM)

* Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
28 4 7 4 2 4 2 2 2 3 6

See also General Properties of Restriction Enzymes:

 

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Updated vasario 06, 2008 13:24