title: Illumina HiSeq 4000 paired end sequencing; Spatial Embryo Profiling of primate gastrulation
design: Spatial Embryo Profiling of primate gastrulation
library_construction_protocol: Cells were dissociated in Accutase for 5 min and pipetted vigorously to obtain a single cell suspension in normal culture media. Single cells were picked by mouth pipette using a blunted microcapillary with internal diameter 15 μM and under a dissection microscope. Cells were collected into 0.2 mL tubes with 4 μL lysis buffer and immediately snap frozen on dry ice. For each sample, RNA separation was performed using biotinylated oligo-dT30VN-tailed oligonucleotides (IDT) conjugated to Dynabeads Streptavidin C1 (65001, Invitrogen) in an RNAse-inhibitor (RNAsin; N2615, Promega) supplemented buffer solution.  Sequencing libraries were prepared according to an adapted version of the G&T protocol previously described. Samples were transferred to a 96-well plate and prepared for both transcriptome and methylome sequencing, by physical separation of genomic DNA (gDNA) from messenger RNA (mRNA).