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16-Apr-2020 01:20

Each 25-μL reaction contained 200 μM of each primer, 200 μM of each d NTP, 2.5 μL 10× buffer (Bioline, Luckenwalde, Germany), 1.5–3.0 mmol Mg Cl region, and reaction mixtures were incubated at 94°C for 3 min, then subjected to 25 cycles of the following profile: denaturation for 30 s at 94°C, annealing for 30 s at 52 or 55°C, and extension for 30 s at 72°C. Products from one individual from each of the two known populations of this species were cloned by using the p GEM–T Easy Vector System (Promega, Madison, Wisconsin, USA) in accordance with the manufacturer's protocol.

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PCR amplifications were performed in a CP2-03 Thermal Cycler (Corbett Research, Mortlake, Australia).This distribution has been cited as evidence of a southern origin of the family (Raven and Axelrod, 1974) has been attributed to Elaeocarpaceae.