Single and half-cell RNA-sequencing in Stentor coeruleus control and beta-tubulin knockdown cells

Stentor coeruleus are giant single-celled ciliates that reach lengths of up to 1 mm. Because of their large size, these cells are easy to manipulate and they also possess the ability to heal wounds and regenerate. These properties are ideal for investigating subcellular pattern formation, as we can physically dissect the cells into anterior and posterior halves. We used single and half-cell RNA-sequencing to assay for changes in transcript localization upon knocking down beta-tubulin, an important component of the microtubule cytoskeleton. , After conducting RNAi knockdown of beta-tubulin in Stentor coeruleus, we bisected the cells using a glass needle. Within 20 minutes of bisection, the whole cells and bisected cells were lysed and prepped for sequencing with the NEBNext Single Cell/Low Input Library Prep Kit for Illumina (Cat. No. E6420). ,