5X Taq PCR Master Mix

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5X Taq PCR Master Mix

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Cat. No. : CMC17
Description : Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activi and a 5´ flap endonuclease activity. Taq 5X Master Mix is an optimized ready-to-use solution containing Taq DNA Polymerase, dNTPs, MgCl2, KCl and stabilizers. It is best for routine PCR applications from templates including pure DNA solutions, bacterial colonies, and cDNA. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA.
Source : An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1.
ReagentsSupplied : MgCl2 (25 mM)
1X Taq Master Mix : 10 mM Tris-HCl (pH 8.6 @ 25°C); 50 mM KCl; 1.5 mM MgCl2; 25 units/ml Taq DNA Polymerase; 0.2 mM dNTPs each; 5% glycerol; 0.08% Ethoxylated tert-octylphenol; 0.05% Tween-20
Applications : PCR; Primer Extension; Microarray Analysis; High-Throughput PCR
Heat Inactivation : No
Unit Definition : One unit of Taq DNA Polymerase is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.
Unit AssayConditions : 1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.
ReactionConditions : 1X Taq Master Mix, DNA template and primers in a total reaction volume of 25 or 50 µl.
GeneralNotes : Taq 5X Master Mix is stable for fifteen freeze-thaw cycles when stored at -20°C. Taq 5X Master Mix is also stable for six months at 4°C, so for frequent use, an aliquot may be kept at 4°C. We recommend assembling all reaction components on ice and quickly transferring the reactions to a thermocycler preheated to the denaturation temperature (95°C).
Reaction Setup :
Component25 µl Reaction50 µl ReactionFinal Concentration
10 µM Forward Primer0.5 µl1 µl0.2 µM (0.05–1 µM)
10 µM Reverse Primer0.5 µl1 µl0.2 µM (0.05–1 µM)
Template DNAvariablevariable<1,000 ng
Taq 5X Master Mix5 µl10 µl1X
Nuclease-free waterto 25 µlto 50 µl
Step
TempTime
Denaturation
95°C30 seconds
30cycles
95°C15-30 seconds
45–68°C15-60 seconds
68°C1 minute per kb
Final extension
68°C5 minutes
Hold4–10°C

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