GSM4223723: Cortical Astrocyte_shPTB_4w-2; Mus musculus; RNA-Seq
The total RNA was purified by a Direct-zol RNA MiniPrep kit (Zymo Research) Total 2ug of total RNA was first converted to cDNA by superscript III first strand synthesis kit with primer Biotin-B-T. The cDNA was purified by PCR Clean-Up Kit column to remove free primers and enzyme. Then terminal transferase was applied to block the terminal of a cDNA 3′ end. Streptavidin-coaged magnetic beads were further applied to isolate cDNAs. After RNA degradation by sodium hydroxide, second-strand cDNA was synthesized by random primer. The second strand cDNA was eluted from beads by heat denaturing. The cDNA was then used as template to construct libraries.