Recently Viewed

Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

PstI

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

FastDigest enzyme available O buffer for 100% activity Optimal incubation at 37°C Not inactivated at 80°C in 20 min Genome qualified LO certified

The PstI restriction enzyme recognizes CTGCA^G sites and cuts best at 37°C in O buffer (Isoschizomers: BspMAI)
  
Loading...
Lambda DNA digested with PstI

Lambda DNA digested with PstI, 0.7% agarose, 28 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

Conditions of high pH, low salt, high glycerol, 8% DMSO can cause star activity (Malyguine, E., et al., Gene, 8, 163-177, 1980). Surrounding sequences: the presence of adjacent runs of G-C base pairs confers significant resistance to cleavage (K. Armstrong, W.R. Bauer. NAR 10, 993-1007, 1982). 100% dUTP incorporation at the recognition site reduces PstI cleavage to 25% (T.C. Glenn et al., Biotechniques. 17, 1086-1090, 1994). PstI will not cut AGCTGCAG when methylated by AluI methyltransferase.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsAlw21I, ApaI, BseSI, Eco24I, Mph1103I, PstI, SacI, SdaI.
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 37°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.
Storage BufferPstI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 200 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.15% Triton X-100, 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
O (Orange) 37°C 50-100 50-100 100 100 50-100 50-100 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0-20 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
28 1 1 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 1 0

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
CTGCA^G
  • Alw21I (BsiHKAI)/FastDigest Alw21I* (GTGCA^C)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)* (GTGCA^C)
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I) (ATGCA^T)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GTGCA^C)
  • HpyCH4V
  • SdaI (SbfI)/FastDigest SbfI (SdaI) (CCTGCA^GG)
  • BfmI (SfcI)/FastDigest SfcI (BfmI)
  • HpyCH4V
  • PstI/FastDigest PstI