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BspPI (AlwI)

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

digested lambda DNA (dam-)
1.4% agarose
     
#ER1321
 Supplied with:
10X Buffer Tango™		 100 u
 
1 ml
#ER1322
Supplied with:
10X Buffer Tango™		 500 u
 
1 ml
Commercial Isoschizomers: AclWI, AlwI

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1321, #ER1322
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 Tango™ 55°C 0.5 80°C 20-50 20-50 0-20 0-20 100 0-20

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 BspPI, approximately 70% of the digested DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. More than 50% of these can be recut.

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

Note
  • Assayed using lambda DNA (dam-) (#SD0021).
  • Incubation at 37°C results in 30% activity.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer Tango™ 100 NR

NR: buffer is not recommended, since enzyme activity is less than 20%.

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
58 0 4 12 10 10 10 10 10 13 4

See also General Properties of Restriction Enzymes:

 

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Updated kovo 05, 2008 16:46