GSM6123638: DAM24.SALINE.P10; Rattus norvegicus; RNA-Seq
2 and 24 hours after receieving either injection, dams were lightly anesthetized with isoflurane in O2, followed by the administration of 0.2 mL of oxytocin (20 USP/mL i.p). Distilled water was used to moisten the teats and milk obtained by gently squeezing its base to manually expel the sample for collection. A microhematocrit tube was used to collect ~20 ul of sample, and samples were stored at -80C until processing. For RNA isolation, milk was spun at 1,000 g to separate the fat layer from the whey. This fat layer was then mixed with an equal volume of PBS and spun again at 3,000 rpm to clean the fat of debris. The miRNeasy Mini Kit (QIAGEN, Cat. #217004) was used to isolate total RNA following the manufacturer's directions. A NanoDrop 2000 spectrophotometer was used to quantify isolated RNA samples, which were then stored at -80C prior to sequencing. A total of at least 190ng of RNA per replicate was prepared for sequencing by The New York Genomic Center using the Tru-Seq Stranded mRNA library preparation kit (Illumina, USA). Five biological replicates were used per experimental group. cDNA libraries were sequenced on an Illumina NovaSeq 6000 in two lanes to obtain 100-bp single-end reads at an approximate sequencing depth of 50 million reads per sample.