title: GSM7623029: CRC Adjacent normal N061; Homo sapiens; OTHER
library_construction_protocol: tissues were lysed in 2% SDS and 25 mM EDTA overnight. Ammonium acetate (10M) was added to make the precipitate of SDS with proteins and was removed by centrifugation at 12,000 rpm. Next, DNA was precipitated using isopropanol, washed with 70% ethanol, and dissolved in dissolved in LTET (Tris 10 mM, EDTA, 0.1 mM + Tween 20,0.1% ) The 16S rRNA gene V3-V4 hypervariable region was amplified using the selected universal primers (Forward 5'-NNNNCCTACGGGNGGCWGCAG-3′, Reverse 5′- GACTACHVGGGTATCTAATCC-3′) that contained overhang nucleotide sequences (Forward 5' TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-[locus specific sequence]-3', Reverse 5'- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-[locus specific sequence]-3') for Illumina barcode indexes.