E12.5 female GCs, RNA-seq #1
Male C57Bl/6J mice homozygous for an Oct4-EGFP transgene were mated to wild-type C57BL/6 females, and gonads were taken from resulting embryos at the indicated time point.  GFP-positive germ cells were isolated by flow cytometry.  Cells were sorted into Trizol (Invitrogen 15596-026) and extracted in 1/10 volume 1-bromo-3-chloropropane (BCP, Sigma B9673).  The aqueous phase was then purified using the RNEasy Micro Kit (Qiagen, 74004) according to the manufacturer’s instructions.  For each sample, 2.84 μl of a 1:10,000 dilution of ERCC RNA Spike-in Mix #1 (Ambion 4456740) was added per 10,000 cells.  Sequencing libraries were prepared using the Illumina Tru-Seq RNA kit (Illumina RS-122-2001).
Whitehead Institute for Biomedical Research
E12.5_female_RNA_1