GSM8486623: Mm, RNAseq, Meis2, E12.5, developing distal forelimb, Rep5; Mus musculus; RNA-Seq
Limbs from mice and bats were dissected in ice cold PBS, snap frozen in liquid nitrogen and stored at -80°C until further processing. Total RNA was extracted using RNeasy Mini Kit (Quiagen, #74106) or RNeasy Micro Kit (Quiagen, #74004) according to the manufacturer's instructions. Limb samples derived from E11.5 and E13.5 as well as CS15 and CS17 embryos were directly homogenized in RTL buffer supplemented with 1% β-Mercaptoethanol and applied to spin columns. Limb tissues from older embryonic stages (E15.5 and CS19) were crushed in liquid nitrogen using Bel-Art SP Scienceware Liquid Nitrogen-Cooled Mini Mortar prior to homogenization. Genomic DNA was removed using the RNase-Free DNase Set (Quiagen, 79254). For gene expression analysis, samples were poly-A enriched and libraries were prepared using the Kapa HyperPrep Kit (Roche, #07962347001). RNA libraries were sequenced on a Novaseq2 with 100 bp paired-end reads. RNA-seq experiments were performed at least in biological duplicates.