alias: Control_1
title: RNA-Seq of Mus musculus: adult male macrophage
design: The first step in the workflow involves purifying the poly-A containing mRNA molecules using poly-T oligo-attached magnetic beads.Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNAfragments are copied into first strand cDNA using reverse transcriptase and random primers. This is followed by second strand cDNAsynthesis using DNA Polymerase I and RNase H. These cDNA fragments then have the addition of a single 'A' base and subsequentligation of the adapter. The products are then purified and enriched with PCR amplification.We then quantified the PCR yield by Qubit and pooled samples together to make a single strand DNA circle (ssDNA circle), which gavethe final library.DNA nanoballs (DNBs) were generated with the ssDNA circle by rolling circle replication (RCR) to enlarge the fluorescent signals at thesequencing process. The DNBs were loaded into the patterned nanoarrays and single-end read of 50 bp were read through on theBGISEQ-500 platform for the following data analysis study. For this step, the BGISEQ-500 platform combines the DNA nanoball-basednanoarrays and stepwise sequencing using Combinational Probe-Anchor Synthesis Sequencing Method.
identifiers.namespace: SUB7275970
identifiers.id: Control_1