Tissue RNA-seq expression data and results of venom LC-MS analysis for Hylyphantes graminicola

Tissue RNA-seq expression data of all genes and results of venom LC-MS analysis from Hylyphantes graminicola (sheet-web spider) for manuscript "Chromosomal-level genome of a sheet-web spider provides insight into the composition and evolution of venom" were saved in this dataset. we sequenced the transcriptomes of different tissues from H. graminicola, including the venom gland, the brain, the silk glands, the abdominal tissue with silk glands removed, and four pairs of legs. A total of 24 samples were used to generate the RNA-seq expression data. Total RNA was extracted using the RNAsimple Total RNA kit (TIANGEN, Beijing, China). PolyA RNA-sequencing (RNA-seq) libraries with an insert size of 250 bp were prepared and sequenced on the Illumina NovaSeq 6000 platform.Pooled sheet-web spider venom samples were reduced with DTT (10 mM, 60 min at 56°C), and alkylated with sufficient iodoacetamide (60 min at room temperature). The protein samples were then concentrated (20 min at 80 V) and separated (90 min at 120 V) in 12% SDS-PAGE gel electrophoresis. The gel was stained by Coomassie Brilliant Blue R-250 and decolored until the bands were visualized clearly. Trypsin treatment was performed at 37°C for 4 h. Peptides were washed three times in 0.1% formic acid (FA) and 3% acetonitrile (ACN) and once in 0.1% FA and 70% ACN. The lyophilized samples were then dissolved in solution A (100% water and 0.1% FA) and injected into a homemade C18 Nano-Trap column (4.5 cm×75 μm, 3 μm) at 55°C. The separated peptides were analyzed by Orbitrap Exploris 480 matched with FAIMS (Thermo Fisher), with an ion source of Nanospray Flex™ (ESI), an ion transport capillary temperature of 320°C, and a spray voltage of 2.1 kV. The compensation voltage of FAIMS was set as -45 and -65, with a scan round time of 1 s, a resolution of 15,000, and an automatic gain control (AGC) target value of 750,000.