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Insects, tissue collection, and RNA-Seq

JL Jean-Marc Lassance
BD Bao-Jian Ding
CL Christer Löfstedt
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Pupae of C. pomonella were purchased from Entomos AG (Switzerland) and were sexed upon arrival to our laboratory. Adult males and females emerged in separate jars in a climate chamber at 23 ± 1 °C under a 16 h:8 h light to dark cycle. On the third day after emergence, pheromone glands of female moths were dissected, immersed in TRIzol Reagent (Thermo Scientific), and stored at − 80 °C until RNA extraction. Two pools of 30 glands were collected 3 h before and after the onset of the photophase, respectively. Total RNA was extracted according to the manufacturer’s instructions. RNA concentration and purity were initially assessed on a NanoDrop2000 (Thermo Fisher Scientific). Library preparation and paired-end Illumina sequencing (2 × 100 bp) were performed by BGI (Hong Kong, PRC). We obtained ~ 66 million QC-passing reads per sample.

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