Illumina HiSeq 2500 paired end sequencing; RNA-Seq  to identify germline stem cell-enriched genes and cystoblast-enriched genes and BMP targeted genes
RNA-Seq  to identify germline stem cell-enriched genes and cystoblast-enriched genes and BMP targeted genes
Ovaries are dissected into 1x PBS on ice and incubated in 5 mg/ml collagenase IV in PBS (Worthington Biochemicals) at RT for 45 min to dissociate the tissue. Cells were washed in PBS and filtered through a 40 μm nylon mesh to remove debris before fluorescence-activated cell sorting (FACS) based on the expression of the germline marker vasa.GFP or differentiation factor bam.GFP using a FACSAria™ Fusion cell sorter (Diva 8 Software; BD Biosciences). Cells were sorted into 1x PBS on ice and RNA was isolated. RNA was isolated using TRIzol (Invitrogen). Total RNA was processed and sequenced. TruSeq® Stranded mRNA assay (Illumina, Inc.) according to the manufacturer's protocol
Experimental Factor: cell type
female germline stem cell
Experimental Factor: phenotype
high BMP expression; constitutively active tkv expression