RNAseq analysis comparing gene expression in Drosophila Gpdh1 mutant and Gpdh1; Ldh double mutants vs control strains

The goal of this study was to determine how loss of Gpdh1 or both Gpdh1 and Ldh influences gene expression in Drosophila larvae. RNA was isolated from Drosopohila Gpdh1 single mutants, Gpdh1; Ldh double mutants, as well as control larvae that were heterozygous for either the Gpdh1 mutations or both Gpdh1 and Ldh mutations. Larvae were collected as described in Li, H., Tennessen, J. M. Preparation of Drosophila Larval Samples for Gas Chromatography-Mass Spectrometry (GC-MS)-based Metabolomics. J. Vis. Exp. (136), e57847, doi:10.3791/57847 (2018). RNA was purified from staged mid-L3 larvae using a RNeasy Mini Kit (Qiagen). Sequencing was performed using an Illumina NextSeq500 platform with 75 bp sequencing module generating 41 bp paired-end reads. After the sequencing run, demultiplexing was performed with bcl2fastq v2.20.0.422.