GSM5114569: Microplast Rep2; Homo sapiens; RNA-Seq
Total RNA was extracted from two biological replicates of microplasts enriched samples using RNeasy mini kit (Qiagen, Hilden, Germany). RNA concentration and integrity were determined with Nanodrop 1000 (Nanodrop, Wilmington, DE,USA) spectrophotometry. Libraries were prepared using the TruSeq RNA sample prep kit V2 as per the manufacturer's protocol (Illumina, San Diego, USA). Briefly, Poly(A) RNA was enriched from total RNA samples using oligo-dT attached beads and ds cDNA was prepared from enriched mRNA samples. cDNA from each sample was attached to a specific index adapter and the cDNA library was further amplified using index adapters. All the libraries were validated using Agilent Technologies2100 Bioanalyzer, normalised and pooled together. 100 base paired end sequencing was performed on Illumina HiSeq 2500 sequencing system at NCBS, Bangalore, India to get a minimum of 30 million reads per library. Paired-end, unstranded.