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PpiI

5'-^ 7(N) G A A C (N)5 C T C (N)13^-3'
3'-^12(N) C T T G (N)5 G A G (N)8 ^-5' 
digested lambda DNA
0.7% agarose
      
#ER1541
 Supplied with:
10X Buffer R
10X Buffer Tango™		 50 u
 
1 ml
1 ml
#ER1542
 Supplied with:
10X Buffer R
10X Buffer Tango™		 250 u
 
1 ml
1 ml
Commercial Isoschizomers: -

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1541, #ER1542
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 R 30°C 0.1 65°C 0-20 0-20 0-20 100 50-100 50-100

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 PpiI, 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
 never overlaps
 never overlaps		 - no effect.
- no effect.
- cleavage impaired.
- no effect.
- no effect.

Note
  • Incubation at 37°C results in 60% activity.
  • PpiI cleaves certain DNA sequences at random 7 or 8 nt away on the top strand of the recognition sequence:
    5'...^7-8(N) G A A C (N)5 C T C (N)13^...3'
    3'...^ 12(N) C T T G (N)5 G A G (N)8^...5'
  • The presence of SAM in a reaction mixture results in incomplete cleavage with PpiI.
  • Greater than 10-fold overdigestion with PpiI may result in star activity.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer R 50-100 50-100

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
5 0 1 2 2 2 3 3 3 2 0

See also General Properties of Restriction Enzymes:

 

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Updated kovo 07, 2008 15:07