title: GSM7074709: Uterus, E85_tdTomato_neg_1; Mus musculus; RNA-Seq
library_construction_protocol: The dissections were executed on ice in cold PBS supplemented with 10% (v/v) fetal bovine serum (FBS; Gemini) using fine-pointed forceps. For each gestational time point, except for E5.5, the uterus wall surrounding the implantation sites was removed. Subsequently, the E6.5 and E7.5 embryos were dissected out of the decidua and digested using 1 mg/ml collagenase type I (Worthington Biochemical) containing 20% (v/v) FBS at 37°C for 5-10 min. The embryos at E8.5, E9.5 and E10.5 were removed and excluded from the study.   Yolk sacs of the embryos at E8.5, E9.5 and E10.5, and the E10.5 placentas were washed in PBS and minced into small pieces of approximately 1-2 mm3 using scissors. The cells were then dissociated at 37°C for 20-30 min using 1 mg/ml collagenase type I (Worthington Biochemical) supplemented with 20% (v/v) FBS.  The decidua of E5.5, E6.5, E7.5, E8.5 and E9.5 embryos were washed in PBS and minced into small pieces of approximately 1-2 mm3 using scissors. The tissues were then repeatedly digested using 0.5% (w/v) collagenase type I (Worthington Biochemical) dissolved in PBS with 5 mM glucose and 20 µl/ml deoxyribonuclease I (Promega) for 10 min and 0.25% trypsin (Gibco) for 5 min at 37°C on a rotator. Trypsin was deactivated using PBS containing 10% (v/v) FBS.  The dissociated cells were filtered through a 40 µm cell strainer (Corning Falcon™, Cat# 352340) and centrifuged at 230 g for 5 min at 4°C. Subsequently, the dissociated cells were washed two times in ice-cold PBS supplemented with 10% (v/v) FBS and resuspended in PBS supplemented with 2% (v/v) FBS. To generate Gel Bead-InEMulsions (GEMs), 12,000 living cells, Chromium Single Cell 3' GEM Kit v3 and Chromium Single Cell 3' Gel Bead Kit v3 were used and loaded on a Chromium Single Cell Controller (10x Genomics). Recovered cDNA was used to construct library using the Single Cell 3' Library kit (10x Genomics), according to the manufacturer's protocol. The libraries were subjected to Illumina HiSeq X Ten and Nova-seq 6000 sequencing system.