title: GSM9151586: Brain Tissue_ Treatment Group_Male 2; Mus musculus; RNA-Seq
library_construction_protocol: Whole brain tissue was rapidly dissected from each mouse, immediately snap-frozen in liquid nitrogen, and stored at -80°C until processing .Total RNA was extracted using Trizol and followed by purfication with the Qiagen RNeasy mini kit to remove contaminants. Non strand specific library Messenger RNAwaspurified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Strand specific library Messenger RNAwaspurified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and and bioanalyzer for size distribution detection.