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Ppu21I (BsaAI)

5'...Y A C↓G T R...3'
3'...R T G↑C A Y...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Unique buffer for 100% activity Optimal incubation at 30°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min Genome qualified LO certified

The Ppu21I (BsaAI) restriction enzyme recognizes YAC^GTR sites and cuts best at 30°C in its own unique buffer (Isoschizomers: BsaAI, BstBAI)
  
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Lambda DNA digested with Ppu21I (BsaAI)

Lambda DNA digested with Ppu21I (BsaAI), 0.7% agarose, 14 cleavage sites

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer Ppu21I: 10 mM Tris-HCl (pH 7.2 at 37°C), 3 mM MgCl2, 150 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 30°C
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
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.
Storage BufferPpu21I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X		 G (green)
1X		 O (orange)
1X		 R (red)
1X		 Tango
(yellow)
1X / 2X
Ppu21I Buffer (Unique) 30°C 50-100† 100† 20-50 NR NR NR NR

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: completely overlaps - blocked.
  • EcoKI: may overlap - effect not determined.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpGYACGTR Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
14 2 5 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 0 2

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
CAC^GTR
  • AjiI (BmgBI)* (CAC^GTC)
  • ZraI (GAC^GTC)
  • AjiI (BmgBI)
  • MaeII
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • AluI/FastDigest AluI (AG^CT)
  • CviJI* (RG^CT)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • SetI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • SetI
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • FaiI* (YA^TG)
  • TscAI (TspRI)/FastDigest TspRI (TscAI)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • BceAI
TAC^GTR
  • AjiI (BmgBI)* (CAC^GTC)
  • ZraI (GAC^GTC)
  • MaeII
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • AluI/FastDigest AluI (AG^CT)
  • CviJI* (RG^CT)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • SetI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • SetI
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • BceAI