title: GSM5425783: Liver_Control_4; Sus scrofa; RNA-Seq
library_construction_protocol: liver biospies were removed, flash frozen on liquid nitrogen, and RNA was harvested using Trizol reagent. RNA from each liver was isolated using Tri-reagent (Ambion). DNA contaminants were removed by TURBO DNAse treatment using the DNA removal kit from Ambion. RNA was quantified by absorbance at A260/280. The integrity, concentration and purity of RNA was verified by a Biorad experion chip (Hercules, CA, USA) 8 liver RNA samples from representative individuals were pooled in 4 pools for each condition. The first step in the workflow involves purifying the poly-A containing mRNA molecules using poly-T oligoattached magnetic beads. Following purification, the mRNA is fragmented into 100 bp pieces. The cleaved RNA fragments are copied into doble strand cDNA.containing the addition of a single 'A' base and the subsequent ligation of the adapter. After purification and enrichment by PCR, samples were pooled to make single strand DNA circle(ssDNA circle), which gave the final library. DNA nanoballs (DNBs) were generated with the ssDNA circle by rolling circle replication (RCR) to enlarge the fluorescent signals at the sequencing process. The DNBs were loaded into the patterned nanoarrays and pair-end reads of 100 bp were read through on the DNBseq platform for the following data analysis study.