Term: design_description

Free-form description of the methods used to create the sequencing library; a brief 'materials and methods' section.

Origin: NCBI

Example: samples were collected and filtered onto Sterivex 0.22 um cartridge filters. DNA was extracted from Sterivex by adding lysis buffer and magnetic bead-based extraction kits (ZymoBIOMICS 96 DNA/RNA MagBead kit). Following bead beating in the cartridges, extractions were finished on an automated KingFisher Flex instrument (Thermo Fisher) in 96-well plates. A two-step PCR approach was used, targeting 16S V4-V5 rRNA with primers 515F (5-GTGYCAGCMGCCGCGGTAA-3) and 926R (5-CCGYCAATTYMTTTRAGTTT-3). Primers were constructed with Fluidigm common oligos CS1 forward (CS1-TS-F: 5-ACACTGACGACATGGTTCTACA-3) and CS2 reverse (CS2-TS-R: 5-TACGGTAGCAGAGACTTGGTCT-3) fused to their 5' ends. PCR products were sent to the Michigan State University Research Technology Support Facility Genomics Core for secondary PCR and sequencing. Secondary PCR used dual-indexed, Illumina-compatible primers, targeting the Fluidigm CS1/CS2 oligomers at the ends of the PCR products. Sequencing runs were performed on an Illumina MiSeq to produce 250+250 nt paired reads.

Sheet(s) containing term: prep_data

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