E12.5_XX_Kit+/Kit+_ovary 1
Total RNA, of about 1 ug, was extracted using Trizol (Invitrogen) according to manufacturer’s instructions. Hemoglobin transcripts were selectively removed from total RNA by following GLOBINclear (Ambion) protocol recommendations. As per the Illumina kit protocol, poly-A-selected mRNA was used to generate mRNA-seq cDNA libraries using random hexamer-primers. cDNA fragments of ~200 nucleotides were isolated and modified for sequencing following the mRNA-seq protocol (Illumina). Illumina Genome Analyzer II was used to sequence 36-mers from the mRNA-seq libraries by following the manufacturer's recommendations.
Whitehead Institute
E12.5_XX_Kit+/Kit+_ovary 1