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[email protected] PRODUCT INFORMATION HyTaq DNA polymerase Kit MB-639200/ 1000U Product information: HyTaq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. The enzyme is isolated from Thermus aquaticus and has a molecular weight of approximately 94 kDa. HyTaq DNA Polymerase has both a 5'→3' DNA polymerase and a 5'→3' exonuclease activity. The enzyme lacks a 3'→5' exonuclease activity (no proofreading ability). Taq DNA Polymerase leaves an A′ overhang, which makes the enzyme ideal for TA cloning. Storage: This product is stable for two years at -20C Unit Definition One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions. General PCR protocol: This protocol serves as a guideline for primer extensions. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined. Notes: Set up reaction mixtures in an area separate from that used for DNA preparation or product analysis. 15 mM MgCl2 is present in the 10X PCR Enhancer buffer. The 1X concentration is 1.5 mM MgCl2. 1. Thaw 10X PCR Enhancer Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts. 2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA. [email protected] Table 1. Reaction components (master mix and template DNA) Component Vol./reaction Final Conc. 10X PCR Enhancer Buffer 5 μL 1X dNTP mix (12.5 mM of each) 0.8 μL 0.2 mM of each dNTP Primer A Variable 0.1–1.0 μM Primer B Variable 0.1–1.0 μM HyTaq DNA Polymerase 0.8 μL 2units Distilled Water Variable ---- Template DNA Variable Variable TOTAL volume 50 μL ---- Related products: dNTP Solution 2x Taq Master Mix RED Nuclease free water 100bp DNA Ladder 1kb DNA Ladder DNAsafe Nuclease Dye Agarose 10X TBE Buffer 50X TAE Buffer