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Ppu21I (BsaAI)
 Available in FastDigest® format

5'-Y A C^G T R-3'
3'-R T G^C A Y-5'
digested lambda DNA
0.7% agarose
         
#ER1971
 Supplied with:
10X Buffer Ppu21I		 500 u
 
1 ml
Commercial Isoschizomers: BsaAI, BstBAI

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1971
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, %
Ppu21I
(unique)		 B 
(blue)		 G 
(green)		 O 
(orange)		 R 
(red)		 Tango™ 
(yellow)
1X 1X 1X 1X 1X 1X 2X
Buffer Ppu21I 30°C 0.5 65°C 100 50-100* 100* 20-50 NR NR NR

NR: buffer is not recommended, because of high star activity.
* Star activity appears at 5-fold overdigestion (5 units x 1 hour).

Storage Buffer
 Ppu21I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 7 mM 2-mercaptoethanol, 0.2 mg/ml BSA and 50% (v/v) glycerol.

Ligation and Recleavage
After 50-fold overdigestion with Ppu21I, more than 90% of the digested DNA fragments can be ligated and recut.

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

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
  • Incubation at 37°C results in less than 30% activity.
  • Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity.

Double Digestions using Tango™ Buffer

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer Ppu21I 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
14 2 5 1 0 0 1 1 1 0 2

See also General Properties of Restriction Enzymes:

 

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Updated balandžio 22, 2008 11:33