Transcriptome of Lordiphosa clarofinis adult head (apple-fed) C4_4 (CRX1973561; CRA029924)
Smart-seq2-based bulk RNA sequencing sequencing. Sampled tissue: adult head; feeding condition: apple (15 days post-feeding). Tissues were dissected, rapidly frozen in liquid nitrogen and stored at -80C. All sampled tissues were washed 5x with PBS in a clean bench, minced, suspended in enzyme mixture (collagenase IV 1 mg/ml + DNase I 0.2 mg/ml), dissociated into single cells at 37C for 15 min (agitated every 5 min), washed 3x, filtered through 40-um strainer. Single cells were washed 3x in 0.1% BSA-PBS droplets, manually picked into lysis buffer (2 uL 0.2% Triton X-100 + 1 uL oligo(dT) primer + 1 uL dNTP) via mouth pipette under microscope. Reverse transcription and 23 cycles of amplification were done to get cDNA. Libraries were constructed with Nextera XT DNA Library Preparation Kit and sequenced on Illumina NovaSeq 6000 (150 bp paired-end). Experimental design: Transcriptome analysis of Lordiphosa clarofinis adult heads under apple-feeding, with 5 biological replicates.