Genomic DNA Extraction from Filters

Genomic DNA was directly extracted from filters as described in Zhou et al. (31 (link)) with the following modifications: all extraction steps were performed with 50 µl proteinase K (10 mg/ml), and after isopropanol precipitation, pelleted nucleic acids were obtained by centrifugation at 50 000g for 30 min at room temperature. The genomic DNA was stored at −20°C until PCR amplification and metagenomic sequencing were carried out.

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Klindworth A., Pruesse E., Schweer T., Peplies J., Quast C., Horn M, & Glöckner F.O. (2012). Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Research, 41(1), e1.

Publication 2012

Centrifugation Genomic Isopropanol Metagenomic Nucleic acids Proteinase k

Corresponding Organization :

Other organizations : University of Vienna, Max Planck Institute for Marine Microbiology, University of Bremen, Constructor University

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Variable analysis

independent variables

Modifications to the extraction protocol described in Zhou et al. (31)

dependent variables

Genomic DNA yield and quality for PCR amplification and metagenomic sequencing

control variables

Proteinase K concentration (50 µl of 10 mg/ml)
Centrifugation conditions (50,000 g for 30 min at room temperature)
Storage of genomic DNA at -20°C

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

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