Taq Polymerase

PowerQ® Taq DNA Polymerase for qPCR

Storage condition: The undiluted enzyme solution is stable when stored at -20℃.

Storage and dilution buffer: 20mM Tris-HCl; 1mM dithiothreitol; 0.1mM EDTA; 0.1M KCl; Nonidet P40, 0.5%(v/v); Tween 20, 0.5%(v/v); glycerol, 50%(v/v); pH 8.0 (4℃).

Concentration:5U/ul

Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10mmols of dNTP into an acid-insoluble material in 30 minutes at 74℃. The reaction conditions are: 50mM Tris-HCl, (pH 9.0 at 25℃), 50mM NaCl, 10Mm MgCl₂, 200uM dATP, dCTP, dGTP and radiolabelled dTTP, and 12.5ug activated calf thymus DNA in a 50ul reaction.

10×Reaction buffer: 100mM Tris-HCl; 500mM KCl; pH 9.0 (20℃). Buffer is optimized for use with 0.2mM for each of dNTPs. A tube of 25mM MgCl₂ is supplied separately.

Note: It is recommended to add dNTPs to this incubation mixture shortly before use. This is to prevent decomposition of the deoxynucleoside thiphosphate that occurs during Prolonged storage at the alkaline pH values required for optimal enzyme activity.

Quality control

Each lot of Taq DNA Polymerase is tested for activity in PCR and efficient incorporation of digoxigenin-11-dUTP, and in DNA sequencing of M13mp18ssDNA. A minimum of 250 bases must be clearly legible in the sequencing gel. Each lot of Taq DNA ploymerase is tested for contaminating activities as stated below.

Test buffer: 10mM Tris-HCl; 1.5mM MgCl₂; 50mM KCl; pH 9.0 (20℃). 0.1mg/ml gelatin.

Absence of endonucleases: 1ul lambda DNA is incubated with Taq Dna polymerase in 50ul test buffer with a paraffin oil overlay at 65℃ for 16 hours. The amount of enzyme showing no degradation of the lambda DNA is ststed under “Endolâ€�.
manual