|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-T^C G A-3'
3'-A G C^T-5'
Thermus aquaticus YT-1
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.
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.
10mM Tris-HCl pH 7.5, 300mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol.
Bsp119I, Bsu15I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, Ssil, XmiI
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.
Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Enzyme is not inactivated by incubation at 80°C for 20min.
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA dam-x 16 hours) with TaqI.
The ligation and recleavage assay was replace with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
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.
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: