Rhagoletis cerasi Larvae
Adults and larvae of Rhagoletis cerasi were flash-frozen in liquid nitrogen and homogenized separately using a sterilized mortar and pestle. Homogenized tissue (100 mg) was transferred to pre-frozen microcentrifuge tubes for RNA extraction. TRI Reagent RNA isolation kit (Sigma-Aldrich) was used following the manufacturer's guidelines. RNA integrity was analyzed using 1.0% agarose gel electrophoresis, and purity and concentration were measured using a Nanodrop spectrophotometer. RNA samples underwent a four-step quality check: agarose gel analysis, RNA purity testing (OD260/OD280) with a Nanodrop, RNA quantification using a Qubit fluorometer, and RNA integrity assessment using an Agilent 2100 instrument. Only samples passing these checks were sequenced using the Illumina Novaseq 6000 platform, generating FASTQ files with sequence and quality information.