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BveI (BspMI)

5'-A C C T G C (N)4^-3'
3'-T G G A C G (N)8^-5' 
digested lambda DNA
0.7% agarose
       
#ER1741
Supplied with:
10X Buffer O
10X Buffer Tango™
50X oligonucleotide		 250 u
 
1 ml
1 ml
25 µl
Commercial Isoschizomers:
Acc36I, BfuAI, BspMI

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1741
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 O
 + 0.5 µM of oligonucleotide		 37°C 0.2 65°C 0-20
(+oligo)		 20-50
(+oligo)		 100
(+oligo)		 20-50
(+oligo)		 50-100
(+oligo)		 100
(+oligo)

Storage Buffer
 10 mM Tris-HCl (pH 7.5 at 25°C), 150 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 50-fold overdigestion with BveI, more than 90% of the digested DNA fragments can be ligated and recut.

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

Note
  • At least two copies of BveI recognition site are required for efficient cleavage.
  • Inclusion of 0.5 µM oligonucleotide with the BveI recognition sequence in the reaction mixture significantly improves cleavage of plasmid DNAs, especially of those with a single BveI site. Still, a complete cleavage of some substrates with BveI is difficult to achieve.
  • BveI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase.
  • Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity.
  • BveI 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 O
 + 0.5 µM of oligonucleotide		 50-100
(+oligo)		 100
(+oligo)

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
41 3 3 1 1 0 1 0 0 0 1

See also General Properties of Restriction Enzymes:

 

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Updated vasario 06, 2008 11:19