GSM4633506: P1 Control female fl/fl Msi1 fl/fl Msi2; Mus musculus; RNA-Seq
Pituitaries from 8-week old control diestrous FVB females (n = 6) were collected and individually dispersed as previously described (14).  Following mechanical dispersion, cells were pelleted and washed once with DMEM.  Cells were pelleted again and fixed with methanol according to the 10x Genomics methanol fixation protocol. Single cell 3' library generation was performed using a 10X Genomics Chromium Controller and the v3 reagent kit (10x Genomics) according to the manufacturer's protocol.  Cell suspensions were loaded on to the middle four channels of a Chromium Single Cell A Chip according to the manufacturer's instructions, aiming for 10,000 cells per channel.  Following generation of single-cell gel bead-in-emulsions (GEM), reverse transcription, fragmentation and cDNA amplification, library preparations and bar coding were performed using the Illumina TruSeq Stranded mRNA Sample Preparation Kit v2 as per manufacturer's instructions 200pM of each library was pooled for sequencing on the Illumina NextSeq 500 platform using a high output flow cell (100 cycles) to generate approximately 25 million 75 base reads per sample.