GSM6258486: reg_batch1_rep228; Mus musculus; RNA-Seq
The fresh isolated retinas at one day post retrograde labeling were dissociated with AMES solution. FACS purification with BD Influx System cell sorter was gated as: Alexa Fluor 488 positive but Texas Red negative RGCs were collected as surRGCs; Texas Red positive RGCs were collected as regRGCs. Individual cell was directly sorted into individual well of 96 well plate with 4ul pre-filled cell lysis buffer, containing 1 U/µl of Recombinant RNase inhibitor (Clontech), 0.1% Triton X-100 (Thermo), 2.5mM dNTP (Thermo), 2.5 µM oligodT30VN (5'AAGCAGTGGTATCAACGCAGAGTACT30VN-3', IDT). Cells were immediately spun down after sorting and frozen at -80°C. RNA libraries were prepared for sequencing using standard Illumina protocols