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Metodología - Coggle Diagram
Metodología
Sample Collection
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Locations:
Rib Reef (18°29'4.8"S, 146°52'13.7"E)
Davies Reef (18°49'23.8"S, 147°38'56.2"E)
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Sampling Period:
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5 time points: February 2014, October/November 2014, February 2015, April 2015, October/November 2015
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PCR and Sequencing
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PCR conditions:
16S PCR:
Volume: 10 µL (triplicate, total 30 µL)
Reagents: AmpliTaq Gold MasterMix, primers, template DNA
Ciclos: 95°C 10 min, 30 ciclos (95°C 30 s, 57°C 60 s, 72°C 60 s), 72°C 7 min
ITS2 PCR:
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Reagents: Qiagen Multiplex MasterMix, Milli Q, primers, template DNA
Ciclos: 95°C 5 min, 31 ciclos (95°C 30 s, 55°C 30 s, 72°C 30 s), 72°C 5 min
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Sequencing and Analysis
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Sequencing:
Illumina MiSeq, 2 bp × 300 bp
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