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PacI

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

FastDigest enzyme available Unique buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 65°C in 20 min Genome qualified Recombinant enzyme LO certified

The PacI restriction enzyme recognizes TTAAT^TAA sites and cuts best at 37°C in its own unique buffer
  
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pJET1 DNA/SdaI fragments with inserted PacI recognition site digested with PacI

pJET1 DNA/SdaI fragments with inserted PacI recognition site digested with PacI, 1% agarose, 2 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
Compatible EndsBstKTI, PvuI.
Conditions for 100% Activity
  • 2X Buffer PacI: 10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10 mM MgCl2, 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 pJET1 DNA with inserted PacI recognition sequence in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferPacI is supplied in: 10 mM potassium phosphate (pH 7.5 at 25°C), 200 mM NaCl, 1 mM DTT, 1 mM EDTA, 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
PacI Buffer (Unique) 37°C 20-50 20-50 0-20 0-20 0-20 0-20 NR

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: may overlap - effect not determined.
  • 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
20-50 50-100

Number of recognition sites in DNA molecules

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

New sites generated by ligation

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

Recognition SequenceSecond EnzymeEnzymes recognizing newly generated recognition sequence
TTAAT^TAA
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)* (CGAT^CG)
  • PvuI/FastDigest PvuI (CGAT^CG)
  • SfaAI (AsiSI)/FastDigest AsiSI (SfaAI) (GCGAT^CGC)
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I

Newly generated recognition sites resulting from removal of 3'-overhang and self-ligation

Recognition SequenceNewly generated sequence after reactionRestriction enzymes that cleave the newly generated recognition sequence
TTAAT^TAATTATAA
  • AanI (PsiI)/FastDigest PsiI (AanI)
  • FaiI