GSM5222800: E14_singlecell_control1; Mus musculus; RNA-Seq
Soft palate tissue (posterior third of the palatal region) was digested from E14.5 wild type embryos by TrypLE express enzyme (Thermo Fisher Scientific) at 37°C with shaking at 600 rpm for 20 min. Full-length cDNA was reverse transcribed from poly-adenylated mRNA using primers containing: an illumina TruSeq Read 1, 16 nt 10x Barcode, 12 nt unique molecular identifier (UMI) and 30 nt poly(dT) sequence. Silane magnetic beads are used to purify the first-strand cDNA from post GEM-RT reaction mixture. Barcoded, full-length cDNA is amplified via PCR to generate sufficient mass for library construction. Enzymatic fragmentation and size selection are used to optimize the cDNA amplicon size. P5, P7, a sample index and TruSeq Read 2 are added via End Repair, A-tailing, Adaptor Ligation, and PCR. The final libraries contain the P5 and P7 primers used in Illumina bridge amplication. Chrominum Single Cell 3' Gene Expression Library
GEO
GSM5222800