Cfr42I (SacII) (10 U/µL)
Detailní popis
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.
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
- Dam: never overlaps - no effect.
- Dcm: never overlaps - no effect.
- CpG: completely overlaps - blocked.
- EcoKI: never overlaps - no effect.
- EcoBI: never overlaps - no effect.
Methylation type | Sequence | Cleavage effect |
---|---|---|
CpG | CCGCGG |
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 |
|
|
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^GG |
CCGG |
|
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