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Cfr42I (SacII)

5'...C C G C↓G G...3'
3'...G G↑C G C C...5'

B buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min Genome qualified Recombinant enzyme LO certified

The Cfr42I (SacII) restriction enzyme recognizes CCGC^GG sites and cuts best at 37°C in B buffer (Isoschizomers: KspI, SacII, Sfr303I, SgrBI, SstII)
  
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Lambda DNA digested with Cfr42I (SacII)

Lambda DNA digested with Cfr42I (SacII), 0.7% agarose, 4 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

Cfr42I activity is affected by high salt concentration. Trace amounts of sodium chloride remaining in the substrate DNA after completion of upstream applications may inhibit enzyme activity and result in impaired DNA cleavage. Certain sites in lambda DNA are difficult to cleave with Cfr42I, the same as with its prototype SacII. At least two copies of Cfr42I recognition site are required for efficient cleavage.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer B: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 20 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 BufferCfr42I 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
B (Blue) 37°C 100 50-100 0-20 0-20 50-100 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
CpGCCGCGG 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
4 1 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 1 1 1 1

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
CCGC^GG
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)* (CGGC^CG)
  • SsiI (AciI)/FastDigest AciI (SsiI)

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

Recognition Sequence Newly generated sequence after reaction Restriction enzymes that cleave the newly generated recognition sequence
CCGC^GGCCGG
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI