GSM4403001: PGR-A:OE replicate 1; Mus musculus; RNA-Seq
Tissues were homogenized by the bead Mill 24 homogenizer (15-340-163, Fisher Scientific, Waltham, MA) in Bead Mill Tubes (15-340-154 Fisher Scientific, Waltham, MA) with 1 mL Trizol (15596026, Thermo Fisher Scientific, Waltham, MA). Tissue debris was pelleted and removed by centrifugation at 12000 xg for 10 minutes at 4°C. After adding 200 uL 1-Bromo-3-chloropropane, samples were manually shaken for 20 seconds followed by incubation at room temperature for 3 minutes. Phase separation was conducted by centrifugation at 12000 xg for 18 minutes at 4°C. The RNA containing aqueous layer was retained and subsequently mixed with 500 uL 200 proof ethanol. The mixture was then passed through the RNA binding column of RNeasy Mini Kit (74104, Qiagen, Germantown, MD), followed by washing and elution steps described in the manufacturer's handbook. RNA libraries were prepared for sequencing using the Illumina TruSeq RNA Library Prep Kit. Paired end, 75bp.
GEO
GSM4403001