RNAseq of heat shocked C. elegans N2
RNA collection was performed on ~800 animals at day 1 of adulthood. Animals were collected into Trizol solution and worms were freeze/thawed 3x with liquid nitrogen with a 30 sec vortexing step between each freeze thaw cycle. After the final thaw, chloroform was added to a 1:5 chloroform/Trizol ratio and aqueous separation was performed using centrifugation in a heavy gel phase-lock tube (VWR, 10847-802). The aqueous phase was mixed 1:1 with isopropanol and then RNA was extracted as per manufacturers directions using a Qiagen RNeasy Mini Kit (74106). Library preparation was performed using a Kapa Biosystems mRNA Hyper Prep Kit as per manufacturers protocol and sequencing was performed at the Vincent J Coates Genomic Sequencing Core at the University of California, Berkeley using an Illumina HS4000 mode SR100.
SUB15921665
LJRH59