title: GSM7159363: RNA-Seq, WT2; Mus musculus; RNA-Seq
library_construction_protocol: 5-10 pseudopregnant D4 WT mouse uterine horns were cut into small pieces (2–3 mm). Tissue pieces were first digested in 5 ml fresh medium (HBSS antibiotic; Gibco) containing 6 mg/ml dispase (Gibco) and 25 mg/ml pancreatin (Sigma), and then incubated in fresh medium (3 ml) containing 0.5 mg/ml collagenase (Sigma) at 37°C for 30 min. The digested cells were passed through a 70-lm filter to obtain the stromal cells. Cells were plated at 60-mm dishes, containing DMEM and Ham F-12 nutrient mixture (1:1) (Gibco) with 10% charcoal-stripped fetal bovine serum (FBS) and antibiotic. After 4 h, the medium was replaced with fresh medium (DMEM/F-12, 1:1) with 10% FBS. 2 hours later, wash twice with PBS gently for the preparation of RNA-Seq samples. The libraries were constructed by using a Universal RNA-Seq Library Preparation Kit (TECAN) per the manufacturer's protocol in which double-strand cDNA was generated using a mixture of random and poly(T) priming, followed by fragmentation of double-stranded cDNA, end repair, adaptor ligation, strand selection, targeted transcript depletion with AnyDeplete, and PCR amplification to produce the final library.