Single-cell RNA sequencing of major ampullate glands from adult female L. sclopetarius spiders
Single-cell RNA sequencing of major ampullate glands from adult female L. sclopetarius spiders
L. sclopetarius adult female spiders were collected in the wild in a small habitat in Uppsala, Sweden. Taxonomic identity of the spider was verified by the Museum of Natural History, Stockholm, Sweden. The spiders were kept in big containers that allowed them to spin webs. They were fed with meal worms or Drosophila flies weekly and watered daily. In total 20 spiders were used for single cell sequencing.  Method A: The major ampullate glands were taken out in ringer solution, pH 7.4. The duct was removed. The glands were washed with PBS, pH 7.4 and then incubated into pre-warmed trypsin-EDTA (Gibco, 0.5%) at 37°C for 1 min in a low-binding micro-centrifuge tube. The glands were triturated with a pipette briefly and centrifuged at 300 g for 3 min at 4°C. The pellets were resuspended in 300 μL DMEM (Gibco) containing 1% BSA (Sigma). The DMEM and the tubes were briefly bubbled with carbogen prior to resuspension. The suspension was strained through a 40 μm cell strainer and transferred to low-binding micro-centrifuge tubes. The library preparation was done using 10X 3' GE kit on the 10X Chromium Single Cell 3' Platform following manufacturer's protocol (Dual Index 10X_3'_V3.1).