NextSeq 500 paired end sequencing; Single cell analysis of emergent haematopoiesis in the human fetal bone marrow (SS2)
Samples were collected by the Human Developmental Biology Resource and subsequently dissected by their team. Single cells suspensions were made by mechanical digestion and live, enriched cells were selected by FACS. Nucleic acid extraction was conducted using an integrated bead-based approach, with cell lysis being a component of the sequencing A slightly modified Smart-seq2 protocol was used. After cDNA generation, libraries were prepared (384 cells per library) using the Illumina Nextera XT kit (Illumina). Index v.2 sets A, B, C and D were used per library to barcode each cell before multiplexing.