Illumina NovaSeq 6000 paired end sequencing; RNA-seq analysis of gene expression in macrophages progenitors from the yolk sac of murine embryos at embryonic days 9 and 10, and the modifications resulting from a defective expression of the transcription factor Lyl-1
RNA-seq analysis of gene expression in macrophages progenitors from the yolk sac of murine embryos at embryonic days 9 and 10, and the modifications resulting from a defective expression of the transcription factor Lyl-1
Macrophage (MF) progenitors (CD45+CD11b+cKit+) were sorted from the yolk sac (YS) of either wild type  (WT) or Lyl-1LacZ/LacZ  (Mut) embryos at embryonic day (E) 9 (primitive MF progenitors only) or at E10 (primitive MF progenitor + MF progenitor derived from erythro-myeloid progenitors).  Total RNA was extracted from sorted cells using Trizol (ThermoFisher). RNA integrity (RNA Integrity Scoreâ‰¥7.0) was checked on the Agilent 2100 Bioanalyzer (Agilent) and quantity was determined using Qubit (Invitrogen). SureSelect Automated Strand Specific RNA Library Preparation Kit was used according to manufacturer's instructions with the Bravo Platform. Briefly, 50ng of total RNA sample was used for poly-A mRNA selection using oligo(dT) beads and subjected to thermal mRNA fragmentation. The fragmented mRNA samples were subjected to cDNA synthesis and further converted into double stranded DNA using the reagents supplied in the kit, and the resulting dsDNA was used for library preparation.