|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 37ēC 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 37ēC.
10mM Tris-HCl pH 7.4,100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periodsthe 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.
AciI, 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.
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.
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.
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: