title: GSM2401366: WTGF_2; Danio rerio; RNA-Seq
library_construction_protocol: Zebrafish digestive tracts (n = 13 – 20 per condition per genotype) were removed by microdissection and resuspended in 1 mL TRIzol (Ambion/Invitrogen/ThermoFisher Scientific 15596026).  Larval digestive tracts were lysed by being passing through a 25 G needle followed by a 27.5 G needle 5 times each.  200 uL of chloroform was added to the TRIzol and the sample was vortexed on high for 30 seconds at room temperature.  The samples were incubated at room temperature for 2 minutes and centrifuged at 12,000 x g for 15 minutes at 4°C.  The top aqueous layer was removed and added to equal volume of isopropanol.  The nucleic acids were isolated using a column-based RNA-isolation kit (Ambion Cat 12183018A) with an on column DNase I (RNase-free) treatment (New England Biolabs M0303L) to remove DNA contamination.  RNA was eluted off the column in nuclease-free water, quantified using a Qubit 2.0 and stored at -80°C until submission to the Duke Sequencing and Genomic Technologies Core. RNA-seq libraries were prepared and sequenced by Duke Sequencing and Genomic Technologies Core on an Illumina HiSeq 2500 with 4 samples per lane in the flow cell.
identifiers.namespace: GEO
identifiers.id: GSM2401366