GSM4336385: Control Rep2; Cochliomyia hominivorax; RNA-Seq
Total RNA and DNA of resistant and control samples were isolated for each individual using TRIzol (InvitrogenTM, Thermo Fisher Scientific, Waltham, Massachusetts, USA) from whole bodies of 142 larvae: 44 and 58 surviving individuals respectively from the first and second replicates of the insecticide treatment, and 40 and 60 control larvae from the control group, first and second replicates, respectively. For RNA samples, DNase I (InvitrogenTM, Thermo Fisher Scientific, Waltham, Massachusetts, USA) was used to remove genomic DNA contamination and mRNA-enriched samples were further purified by using Nucleospin RNA Clean-up columns (Macherey & Nagel, DÃ¼ren, Germany). Quantification was performed using the fluorometer Qubit Quantitation Platform (InvitrogenTM, Thermo Fisher Scientific, Waltham, Massachusetts, USA). RNA from single individuals of each replicate was pooled for sequencing. The sequencing was outsourced to the University of Veterinary Medicine, Vienna, Austria and a second biological replicate to LaboratÃ³rio MultiusuÃ¡rios Centralizado, ESALQ-USP (Piracicaba, Brazil). The RNA fragments resulting from the random breaking of the transcripts were converted into a cDNA library using the mRNA-Seq Sample Prep Kit (Illumina, San Diego, USA). The samples were sequenced using the HiSeq 2000 (Illumina, San Diego, USA).
GEO Accession
GSM4336385