| 5'-T^C G A-3' |
|
| 3'-A G C^T-5' |
|
| Catalog # | T1017 |
| Concentration | 10u/ul |
| Source | Thermus aquaticus YT-1 |
| Supplied wtih | R1625-80 Restriction Enzyme Buffer for TaqI, 10X: Supplied as a liquid in |
| 10mM Tris-HCl pH 8.0, 5mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Use as 1x. Incubate at 65°C under paraffin oil in a capped vial. Incubation at 37ºC results in 10% activity. |
| R1625 | Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 at 37ºC. |
| Diluent Buffer | 10mM Tris-HCl pH 7.4,100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used. |
| Storage Buffer | 10mM Tris-HCl pH 7.5, 300mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol. |
| Compatible Ends | AciI, Bsp119I, Bsu15I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, Ssil, XmiI |
| Unit Definition | One unit is defined as the amount of TaqI to digest 1ug of lambda DNA dam- in 1 hour at 65°C in 50ul of assay buffer. |
| Thermal Inactivation | |
| Enzyme is not inactivated by incubation at 80°C for 20min. |
|
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA dam-x 16 hours) with TaqI. |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with TaqI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1.2uM. More than 95% of these can be recut. |
|
| Labeled Oligonucleotide (LO) Assay | |
| No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of TaqI for 4 hours. |
|
|
| Methylation Effects | |
| Dam | May overlap- blocked |
| Dcm, EcoKI, EcoBl | Never overlaps-no effect |
| CpG | Completely overlaps- no effect |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.3 units of TaqI is required for complete digestion of 1ug of lambda DNA in 16 hours at 65°C. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 121 |
| PhiX174 | 10 |
| M13mp18/19 | 12 |
| pBR322 | 7 |
| pUC18/19 | 4 |
| pUC57 | 4 |
| pTZ19R/U | 5 |
