GSM1312925: 3-D shPAX6 LSC-2; Homo sapiens; RNA-Seq
Total RNA was purified by a Picropure RNA isolation kit Briefly, 600ng 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 NucleoSpin Gel and PCR Clean-Up Kit column (Clontech) to remove free primers and enzyme. Then terminal transferase (NEB) was applied to block the terminal of a cDNA 3’end.  Streptavidin-coaged magnetic beads (Life Technology) were further applied to isolate cDNAs.  After RNA degradation by sodium hydroxide, second strand cDNA was synthesized by random priming with primer A-N8. The second strand cDNA was eluted from beads by heat denaturing.  The cDNA was then used as template to construct libraries by amplifies with barcode primers and primer PB. The sequencing was done on Hiseq 2000 system.
GEO Accession
GSM1312925