GSM8150282: CSN48-2_03_K04; Danio rerio; RNA-Seq
The cells are sorted with an Aria III FACS into a 384 well plate pre-loaded with 0.5 µl freshly prepared lysis buffer (0.2 % Tween-20 solution with 4 U/µl murine RNase inhibitor (NEB)). After the sort, the plate is shortly centrifuged and lysed cells stored at -80°C.After thawing the plate, the cells are immediately mixed with 0.5 µl of a primer mix containing 5 mM dNTP (Invitrogen), 0.5 µM dT-primer*, 2 U RNase Inhibitor (NEB), ERCC spike in mix with a final dilution of 1:2*107. RNA is denatured for 3 minutes at 72 °C, followed by reverse transcription at 42 °C for 90 min after filling up to 1.5 µ