Alw21 I (BsiHKAI)
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
For methylation sensitivity refer to product specifications.
Compatible Ends |
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Conditions for 100% Activity |
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Double Digestion | Perform double digestion using DoubleDigest. |
Hazardous | No |
Isoschizomers | Search for commercial isoschizomers using REsearch. |
Storage Buffer | Alw21I is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA and 50% (v/v) glycerol. |
Storage Condition | -20 C |
Reaction conditions
Recommended buffer for 100% activity | Optimal temperature | Enzyme activity in Thermo Scientific buffers, % | Tango buffer for double digestion | |||||
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B (blue )1X |
G (green )1X |
O (orange) 1X |
R (red) 1X |
Tango (yellow) 1X / 2X |
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O (Orange) | 37°C | 0-20 | 20-50 | 100 | 50-100 | 20-50 | 50-100 | 1X or 2X |
Methylation Effects
- Dam: never overlaps - no effect
- Dcm: never overlaps - no effect
- CpG: may overlap - no effect
- EcoKI: may overlap - no effect
- EcoBI: may overlap - effect not determined
Methylation type | Sequence | Cleavage effect |
---|---|---|
CpG | 5'...m5C GWGCWm5C G ...3'3'... Gm5CWCGW Gm5C ...5' |
No effect |
EcoKI (AAC(N)6GTGC ) |
5'...Am6AC(N)6G TGCWC ...3'3'... T TG(N)6Cm6ACGWG ...5' |
No effect |
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 |
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28 | 3 | 3 | 8 | 5 | 5 |
pTZ19R/U | pTZ57R | pBluescriptIIKS(±) | pBluescriptIISK(±) | pACYC177 | pACYC184 |
4 | 4 | 4 | 4 | 2 | 5 |
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 |
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GAGCA^C |
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GAGCT^C |
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GTGCA^C |
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GTGCT^C |
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