title: Scraping-derived Organoid RNA from Subject hT326
design: For RNA-seq, library preparation was performed using the Kapa mRNA Hyper Prep Kit. The protocol involved poly(A) selection of mRNA from 100 ng of purified total RNA from PDOs in 50 µL DNase-free water, followed by fragmentation at 85°C for 6 minutes to achieve 300-400 bp fragments. First- and second-strand cDNA synthesis generated double-stranded cDNA, which underwent end repair, A-tailing, and adapter ligation. Libraries were amplified with 12 PCR cycles and purified to ensure high quality. QC of the libraries included Qubit and Tape Station assessments. Libraries were pooled, and final quantification was performed using Qubit and Tape Station. The RNA-seq pool was sequenced on one F3 flowcell of a Singular G4 PE100.
library_name: hT326-S-PDO-RNA