GSM8695021: raw264.7,Tppu-2; Mus musculus; RNA-Seq
To extract RNA from samples, the Trizol method was used. After successful extraction, the RNA was dissolved in 50 µL of DEPC-treated water. Then, the total RNA was identified and quantified using a Qubit fluorometer and a Qsep400 high-throughput fragment analyzer. Library construction involves enriching polyA-tailed mRNA using Oligo(dT) beads, fragmenting the mRNA, and synthesizing first strand cDNA with random hexamer primers. The second strand is synthesized with dUTP for strand specificity. After ligating adapters, the library is amplified via PCR and purified. Finally, concentration is measured with Qubit, fragment size analyzed with Qsep400, and effective concentration quantified by qPCR.