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PagI (BspHI)
Available in FastDigest® format 

5'-T^C A T G A-3'
3'-A G T A C^T-5'

digested lambda DNA
0.7% agarose
        
#ER1281
Supplied with:
10X Buffer O		 400 u
 
1 ml
#ER1282
Supplied with:
10X Buffer O		 2000 u
 
1 ml
Commercial Isoschizomers: BspHIm, RcaI
"m" indicates different sensitivity to methylation.

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1281, #ER1282
MSDS (English)
MSDS (English-USA)
MSDS (German)
Chart (in pdf, 721 KB)
Brochure (in pdf, 316 KB)

Concentration
10 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 37°C 0.2 80°C 0-20 50-100 100 NR NR NR

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

Storage Buffer
10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 50-fold overdigestion with PagI, more than 95% of the digested DNA fragments can be ligated and recut.

Methylation Effects
Dam:
Dcm:
CpG:
EcoKI:
EcoBI:		  may overlap
 never overlaps
 never overlaps
 never overlaps
 may overlap		 - cleavage impaired.
- no effect.
- no effect.
- no effect.
- cleavage impaired.

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
Low salt, high glycerol (>5%) concentrations, pH >8.0 or large excess of the enzyme may result in star activity.

Double Digestions using Tango™ Buffer

Optimal
buffer

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

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
8 3 1 4 3 3 2 2 2 4 1

See also General Properties of Restriction Enzymes:

 

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Updated balandžio 22, 2008 11:33