Hotstart Taq DNA Polymerase for qPCR

Description:
ShineGene’s hotStart Taq DNA Polymerase is a chemically modified recombinant Taq DNA polymerase. The enzyme is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer dimers and providing higher specificity of DNA amplification. The functional activity of the enzyme is restored during a 15-minutes incubation at 95°C. The activated enzyme maintains the same functionality as Taq DNA polymerase.

10×Reaction buffer(with MgCl2) : 200mM Tris-HCl; 200mM KCl; 15mM MgCl2,pH8.3 (25℃). Buffer is optimized for use with 0.2mM for each of dNTPs.

Hotstart Program:95℃ 2-4min

Concentration: 5U/ul

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

Applications

·     Hot Start PCR

·    qPCR

·    Routine PCR

·    High throughput PCR

·    Multiplex PCR

·    Genotyping

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°C).

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°C. The reaction conditions are: 50mM Tris-HCl, (pH 9.0 at 25°C), 50mM NaCl, 10Mm MgCl2, 200uM dATP, dCTP, dGTP and radiolabelled dTTP, and 12.5ug activated calf thymus DNA in a 50ul reaction.

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.

Standard Protocol

Reaction setup:

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°C). 0.1mg/ml gelatin.

for 16 hours. The amount of enzyme showing no degradation of the lambda DNA is ststed under "Endol".
Hotstart Taq Polymerase manual

Related Link
Peptide Synthesis
Chromas
Make Antibody
Lab Consumables
Gene Synthesis
dNTP