GSM5293419: K8Y HS LY6D+ cells_E3; Mus musculus; RNA-Seq
Prostate was removed and dissected, cells were FACS-isolated based on YFP expression and cell surface markers, and RNA was harvested using RNA/DNA purification micro kit (Norgen, #50300). RNA Sequence library prep was performed according to a modified Smartseq2 protocol (Picelli, 2014) using Superscript II (Thermo Fisher #18064022) for reverse transcription of polyA-RNA and Kapa Hifi Hotstart Readymix PCR kit (#KR0370) for second strand synthesis and amplification. Tagmentation reactions were carried out using a Nextera XT DNA Library Preparation Kit (Illumina, #FC-131-1024). Libraries were quantified using Kapa Library Quantification Kit for Illumina Sequencing Platforms (Roche, #07960336001). Single read 75bp sequencing was carried out by clustering 2pM of pooled libraries on a Nextseq500 sequencer (Illumina). RNA libraries were prepared for sequencing using standard Illumina protocols