GSM5020922: Purified Amyloid-bodies following 60min heat shock 2; Homo sapiens; RNA-Seq
Plates were washed with PBS and cells were harvested in cold Solution 1 (0.5M sucrose, 3mM MgCl2) and sonicated for 6 × 10s using a microtip probe. Samples were then underlaid with cold Solution 2 (1M sucrose, 3mM MgCl2) and centrifuged at 2800 × g for 10 min at 4°C. Pellets were resuspended in PBS and a fraction of each sample was used to determine proper isolation of nucleoli and Amyloid-bodies by microscopic evaluation. Remaining resuspended samples were then used for RNA isolation with Trizol reagent (Invitrogen). Library construction for mRNA sequencing was performed by the SCCC Onco-Genomics Share Resource using the KAPA Stranded mRNA-Seq Kit (KAPA Biosystems).