RNA-seq of Ammoxenus amphalodes venom glands
Paired venom glands were dissected from Ammoxenus spiders that had been milked five days prior to dissection to stimulate venom-related transcript production. To reduce contamination, the dissected glands were immediately immersed in a drop of physiological solution (0.9% NaCl) for 5 min to remove haemolymph, epithelial cells, and other tissues on the surface of the glands. Next, the glands were placed into 2 mL of TRIzol Reagent (Life Technologies, USA) in an Eppendorf tube, compressed with tweezers, and incubated at room temperature for 15 min. The tube was then stored at 20C until further processing. RNA was extracted using TriReagent (MRC Holland, Amsterdam, Netherlands) and RNAeasy spin columns (Qiagen, Hilden, Germany). The tissue was lysed in TriReagent, and RNA was separated into the aqueous phase with 1-bromo-3-chloropropane (MRC Holland, Amsterdam, Netherlands). The aqueous phase was then mixed in a 1:1 ratio with 70% ethanol and transferred to an RNAeasy column. RNA extraction continued according to the manufacturer's protocol, including an on-column DNase digestion.Sequencing of the library was performed on an Illumina HiSeq platform.