TaqI
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
Assayed using Lambda DNA (dam-) (#SD0021). TaqI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163. Incubation at 37°C results in 10% activity. We recommend using TaqI, HC at 37°C.
For methylation sensitivity refer to product specifications.
| Compatible Ends | Bsp119I, Bsu15I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, SsiI, XmiI. |
| Conditions for 100% Activity |
|
| Double Digestion | Perform double digestion using DoubleDigest. |
| Hazardous | No |
| Isoschizomers | Search for commercial isoschizomers using REsearch. |
| Storage Buffer | TaqI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5 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 | |||||
|---|---|---|---|---|---|---|---|---|
| B (blue) 1X |
G (green) 1X |
O (orange) 1X |
R (red) 1X |
Tango (yellow) 1X / 2X |
||||
| TaqI Buffer (Unique) | 65°C | 0-20 | 20-50 | 20-50 | 20-50 | 20-50 | 20-50 | 1X or 2X |
Methylation Effects
- Dam: may overlap - blocked.
- Dcm: never overlaps - no effect.
- CpG: completely overlaps - no effect.
- EcoKI: never overlaps - no effect.
- EcoBI: never overlaps - no effect.
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
Dam(GATC) |
5'...TCGm6A TC...3'3'... AGC Tm6AG...5' |
Blocked |
| CpG | TCGA |
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 | 50-100 | ||
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 121 | 10 | 12 | 7 | 4 | 4 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
| 5 | 5 | 7 | 7 | 10 | 10 |
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 |
|---|---|---|
T^CGA |
|
|
Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation
| Recognition Sequence | Newly generated sequence after reaction | Enzymes that cleave the newly generated sequence |
|---|---|---|
T^CGA |
TCGCGA |
|
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