title: GSM5979812: GD9_4hr_EtOH_Rep4; Mus musculus; RNA-Seq
library_construction_protocol: Either 2 or 4 hr after the injection, dams were sacrificed via CO2 followed by cervical dislocation. Embryos were dissected from the placenta and stage-matched based on somite number (2 hr: 19-20 somites; 4 hr: 20-21 somites). A maximum of two embryos per litter were used to minimize litter effects. RNA from the rostroventral neural tube (RVNT) from each embryo was isolated using the RNeasy Plus Micro kit (Qiagen, Germantown, Maryland) as previously described (Boschen et al., 2020). RNA from a total of 6 embryos per condition and time point (24 total embryos) were submitted for RNA-seq. Libraries for RNA sequencing (RNA-seq) were prepared using the SMARTr Ultra Low Input RNA (Clontech, Mountainview, CA) and Nextera XT DNA (Illumina, San Diego, CA) kits by the UNC High Throughput Sequencing Facility. Samples were pooled prior to sequencing, after RNA extraction and library preparation was complete. Six total pools (1 sample/group/pool) and 1 pool per lane were used. Paired-end (50bp) sequencing was performed on an Illumina HiSeq 4000.