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BplI

5'-^ 8(N) G A G (N)5 C T C (N)13^-3'
3'-^13(N) C T C (N)5 G A G (N) 8^-5'

digested lambda DNA
0.7% agarose
       
#ER1311
Supplied with:
10X Buffer Tango™
50X SAM		 100 u
 
1 ml
0.1 ml
#ER1312
Supplied with:
10X Buffer Tango™
50X SAM		 500 u
 
1 ml
2x0.1 ml
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1311, #ER1312
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 Tango™ + 
0.05 mM S-adenosylmethionine		 37°C 0.3 65°C 0-20
(+SAM)		 20-50
(+SAM)		 0-20
(+SAM)		 0-20
(+SAM)		 100
(+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 Recleavage
After 10-fold overdigestion with BplI, 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 restriction 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 10 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.

Note
  • BplI reguires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05 mM S-adenosylmethionine gives more than a 100-fold increase of BplI activity. Still, a complete cleavage of some substrates with BplI is difficult to achieve.
  • BplI 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 Tango™ + 
0.05 mM S-adenosylmethionine		 100
(+SAM)		 20-50
(+SAM)

Number of Recognition Sites in DNA Molecules

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

See also General Properties of Restriction Enzymes:

 

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Updated kovo 06, 2008 13:30