Tth DNA Polymerase (C09-1068-123)

Product manualTth DNA polymerase is a thermostable enzyme with a molecular weight of 94kDa. It is recombinantly expressed in E. coli from the source Thermus thermophilus HB-8 gene. The enzyme can replicate DNA at 74°C and has a half-life of 20 minutes at 95°C. Tth DNA polymerase can catalyze the polymerization reaction of nucleotides along the 5`→3` direction in the presence of magnesium ions to form double-stranded DNA, or in the presence of manganese ions, use RNA as a template along 5`→ Nucleotide polymerization takes place in the 3'direction. The enzyme also has 5`→3` exonuclease activity. Tth DNA polymerase can be used for PCR, RT-PCR, reverse transcription, and primer extension reactions under higher temperature conditions.Product contentTth DNA Polymerase (5U/μl)2×Tth One Step RT-PCR Buffer (Mg2+ Plus) Mn2+(50×)Storage buffer10mM Tris-HCl (pH 7.5, 25℃), 300mM KCl, 1mM DTT, 0.1mM EDTA, 50% glycerolApplication1）RT-PCR2）PCRInstructions1. System configuration (50μl system)ComponentVolumeFinal concentrationTth DNA Polymerase（5U/μl）1μl—2×Tth One Step RT-PCR Buffer25μl1×Mn2+(50×)1μl1×RNasin（40U/μl）1μl—Primer-probe Mix3μl ※—Template5μl ※—Ultra-pure waterUp to 50μl—Total capacity50μl—※ Note: The amount of primers, probes and templates can be adjusted by yourself. If the system is not enough, please use ultrapure water to make up.2. Reaction conditionsStepTemperatureTimeNumber of cycles195℃2.5min1260℃30min1395℃2.5min1394℃15s 40※455℃40s（Collect fluorescence）. Specification: 5U/μl