title: GSM7163395: E9_AGM_CD31_280218_9; Mus musculus; RNA-Seq
library_construction_protocol: Endothelial and hematopoietic cells were isolated from E8.5-E9.5 yolk sac, E9.5-E11.5 AGM, E11.5-E13.5 placenta and E12.5-E14.5 fetal liver. For AGM we used the method described by (Boisset et al., 2010), in which the head and the tail of the embryo are cut and a buffer containing antibodies (CD31, Kit) is injected into the aorta allowing the simultaneous removal of blood and staining of the inner wall of the aorta. After staining, the AGM was dissected from the trunk. The corresponding tissues were pooled together for further processing. For single-cell suspension, yolk sac was digested with TrypLE for 5-10 min at 37˚C, while AGM and minced placenta were digested in 1.2 mg/ml collagenase I at 37˚C for 1-1.5h. Fetal livers were either grinded or digested with 1.2 mg/ml or 3 mg/ml collagenase I for 5-10 min at 37˚C. The samples were then filtered (YS, placenta = 40 µm; AGM = 35 µm; fetal liver = 30, 40 and 70 µm). Contaminating red blood cells were removed from the single-cell suspension of placenta and fetal liver samples by density gradient centrifugation (Ficoll). As described in CEL-Seq2 protocol (Hashimshony et al. 2016) and its modified version (Herman et al., 2018). Adapted from TruSeq Small RNA Library Preparation Protocol