GSM6505230: RNAseq E10.5 PA3 wt  1; Mus musculus; RNA-Seq
Embryonic tissues were micro-dissected at E10.5, E12.5 or E14.5. Dissected tissue from E10.5 and E12.5 embryos was kept in 1× PBS on ice, then treated with 0.5% trypsin/1× EDTA at 37 °C for 10 minutes and immediately put on ice. Dissected tissue from E14.5 embryos was kept in 1× PBS on ice, then treated with papain digestion mix (10 mg ml−1 papain, 2.5 mM cysteine, 10 mM HEPES (pH 7.4), 0.5 mM EDTA and 0.9× DMEM) for 7 min at 37 °C and immediately put on ice. The tissue was rinsed once in ice-cold 1× DMEM/10% FBS, followed by two washes in ice-cold 1× DMEM. Tissue was dissociated by pipetting. Cranial NCCs from embryos with genotype Wnt1::Cre;ROSARFP were filtered and collected by FACS. After sorting, cranial NCCs were pelleted by centrifugation for 5 minutes, 1000 rpm at 4°C. Total RNA was extracted using the Single Cell RNA Purification Kit (NORGEN, 51800) with genomic DNA digestion using an RNase-Free DNase I Kit (NORGEN, 25710) according to the manufacturers' protocol. For each sample, three independent biological replicates were prepared.  For sequencing of total RNA, unstranded RNA-Seq libraries were prepared using Illumina TruSeq RNA Library Prep Kit v2 according to manufacturers' instructions. Sequencing was performed on an Illumina HiSeq 2500 machine (50 bp read length, single-end)  or an Illumina NovaSeq machine (50bp read length, paired-end). IlluminaTruSeqTotalRNA