GSM3617958: 30348: microbial 16S rRNA genes; human metagenome; OTHER
Genomic DNA was isolated from the respiratory samples with the PowerSoil DNA Isolation Kit The 16S rRNA gene V4 region was amplified with methods published in Caporaso, et al. (2012). The primers included the Illumina flowcell adapter sequences and region-specific sequences, where the reverse primers included barcode sequences. The PCR reaction was carried out using Q5 Hot Start High-Fidelity DNA Polymerase and the conditions were 94 degrees C for 2 min, 33 cycles of 94 degrees C for 30s, 55 degrees C for 30s and 72 degrees C for 1 min 30s, followed by 72 degrees C for 10s and hold at 10 degrees C. The PCR reactions were purified using 0.65X volumes of Agencourt RNAClean XP beads. The library was sequenced using an Illumina PE 2x250 V2 kit on an Illumina MiSeq Sequencer .
GEO Accession
GSM3617958