GSM7098075: Kir21_VIP_Trans_BioRep_5; Mus musculus; RNA-Seq
A nuclear suspension was prepared from frozen visual cortices and subsequently nuclei were FACS sorted directly into a lysis buffer. Libraries were prepared based on Meso-Seq protocol. Briefly, 50-100 nuclei were collected by FACS directly into 2.03 µl lysis buffer (0.2% Triton X-100 in ddW) supplemented with dNTPs (2mM each), SMART-dT30VN primers (1mM), ERCC (1:1.25x107), and RNAse inhibitor. RT was performed directly on isolated nuclei, followed by PCR-amplification (17 cycles). RNA is cleaned using Agencourt Ampure XP beads (Beckman Coulter) and cDNA libraries were generated using Nextera XT (Illumina).