alias: Nestmate_2
title: RNA-Seq of Temnothorax nylanderi: abdomen of healthy adult worker from nest containing infected workers rep2
design: Ant colonies were collected in October 2017 and 2018 from the Lenneberg forest near Mainz, Germany. Ants were transported to the Johannes Gutenberg University Mainz, relocated to slide nests composed of a Plexiglas cavity sandwiched between two microscope slides, and placed in three-chambered nesting boxes. Until sampling, ants were kept at 22C and fed with honey twice weekly but no crickets to prevent contamination of the gut content. Water was provided ad libitum. In November 2017, we selected four infected colonies and transferred four infected and four non-infected workers from the brood-pile per colony into small Petri dishes. Infected workers are mostly found directly on the brood although they show less brood care activity (Scharf et al., 2012). Infection status was preliminarily assessed by the color of the cuticle since infected workers have a yellow cuticle compared to the typical brown cuticle of their uninfected nestmates (Trabalon et al., 2000). However, note that in infected colonies, sometimes infected workers occur with a normal brown cuticle, and without dissection, infection status cannot be determined with certainty (Scharf et al., 2012). We, therefore, confirmed infection status later by mapping the reads of each sample against the cestode transcriptome. The gaster of the ants was detached with a razor blade directly in front of the postpetiole and four abdomens per colony were pooled into 100 l of Trizol to reduce variance between samples. Abdomens were directly crushed in Trizol, which was kept on ice and protected from direct light. Samples were stored under -80C until RNA extraction, which was performed using the RNAeasy mini extraction kit (Qiagen) following the standard protocol and sent to the Institute of Molecular Biology Mainz (IMB) for sequencing on an Illumina NextSeq500. The entire procedure was repeated exactly one year later, in 2018, only this time we selected four uninfected colonies and obtained one pool of four abdomens of again inside workers directly on the brood and one sample of the abdomen of the queen per colony. Thus, the location inside the nest was the same for all our four groups.
identifiers.namespace: SUB8418782
identifiers.id: Nestmate_2