GSM4632649: Ventral foregut endoderm 24_23_rep3; Amia calva; RNA-Seq
Using a cryostat (Leica CM 1950), embedded bowfin embryos were sectioned transversely at 15 µm from the anterior to posterior ends of the gas bladder bud and mounted on polyethylene naphthalate (PEN) membrane slides.From each transverse section, the dorsal and ventral regions of the foregut endoderm and mesoderm were excised using the Zeiss Palm Microbeam LCM System at the Cornell University Biotechnology Resource Center. LCM was completed in 2 hours or less for each slide to minimize RNA degradation in the samples. Immediately following LCM, we stabilized the RNA with buffer RLT with β-Mercaptoethanol (Qiagen RNeasy Micro) and homogenized the sample by vortexing for 2 minutes. RNA extraction was performed using the Qiagen RNeasy Micro Kit. RNA amplification and cDNA reverse transcription was completed using the NuGEN Ovation RNA-seq System V2. We used NEBNext Ultra II DNA Library Prep Kit to prepare cDNA libraries for sequencing. All libraries were quality checked on a Tape Station by the Cornell BRC Genomics Facility and showed a single peak of between 170 and 240 base pairs.
GEO Accession
GSM4632649