GSM2754173: 6_somites_Mutant_rep2; Danio rerio; RNA-Seq
The tip of the tail and a little bit of the yolk close to the tail was dissected using iridectomy scissors at 6 ss to be utilized for genotyping and the rest of the embryo was placed in Trizol Reagent (Life Technologies, Carlsbad, CA). Total RNA was extracted from 6 ss (12 hpf) tbx1 null and wild type embryos using SV Total RNA Isolation System (Promega, Madison, WI). RNA integrity and concentration were checked on a Fragment Analyzer (Advanced Analytical). The mRNA was purified by polyA-tail enrichment, fragmented, and reverse transcribed into cDNA (Illumina TruSeq). cDNA samples were then end-repaired and adaptor-ligated using the SPRI-works Fragment Library System I (Beckman Coulter Genomics) and indexed during amplification. Libraries were quantified using the Fragment Analyzer (Advanced Analytical) and qPCR before being loaded for paired-end sequencing using the Illumina HiSeq 2000. polyA-based mRNA capture using SPRI beads
GEO Accession
GSM2754173