Microbial Community DNA Extraction and Sequencing
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Corresponding Organization :
Other organizations : University of Calgary, Marine Biological Laboratory, Sun Yat-sen University, Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou), Hodges University
Protocol cited in 2 other protocols
Variable analysis
- Homogenization and cell lysis conditions (bead beating at 6 m s^-1 for 45 s)
- Bacterial 16S rRNA gene (v3-4 region) abundance and composition
- Archaeal 16S rRNA gene (v4-5 region) abundance and composition
- Amount of sediment used for DNA extraction (0.5 to 1.0 g)
- DNA extraction method (DNeasy PowerLyzer PowerSoil Kit)
- PCR primer pairs (SD-Bact-0341-bS17/SD-Bact-0785-aA21 for bacteria, SD-Arch-0519-aS15/SD-Arch-0911-aA20 for archaea)
- Sequencing platform (Illumina MiSeq benchtop sequencer)
- Extraction blanks to assess laboratory contamination
- PCR reagent blanks to confirm negative amplification
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