title: Illumina HiSeq 4000 paired end sequencing; Single-cell RNA sequencing of marmoset embryonic development
design: Single-cell RNA sequencing of marmoset embryonic development
library_construction_protocol: Marmoset embryos were collected by non-surgical uterine flush and staged according to cell number and embryonic day. Where appropriate, zona pellucidae were removed using acid Tyrode's solution (Sigma) and embryos were subjected to immunosurgery as previously described (Solter and Knowles, Proc Natl Acad Sci USA 72:5099-102; Nichols et al. Cell 95:379-91) using a custom rabbit polyclonal anti-marmoset antibody (Boroviak et al. Dev Cell 35:366-82). Following the complement reaction, residual trophectoderm was thoroughly removed by repetitive manual pipetting. For dissociation of marmoset ICM into single cells, recovered ICM were exposed to a 1:1 mixture of 0.025% trypsin plus EDTA (Invitrogen) and 0.025% trypsin (Invitrogen) plus 1% chick serum (Sigma) for 5-10min. Cells were dissociated into singletons by repetitive pipetting with glass microcapillaries of gradually reduced inner diameter.  Individual cells were transferred to lysis buffer and snap frozen on dry ice. Single-cell RNA-seq libraries were prepared according to the Smart-seq2 protocol (Picelli et al. Nat Methods 10:1096-8; Picelli et al. Nat Protoc. 9:171-81). No spike ins were used for this dataset.
attributes.tag: Experimental Factor: single cell identifier
attributes.value: LateICM_4_C19
attributes.tag: Experimental Factor: developmental stage
attributes.value: late blastocyst