title: GSM8902288: BY4741_SDS_8hr_bio_rep2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: We extracted total RNA using an RNeasy mini kit (QIAGEN, Germany). We added 600 µl of RTL buffer and glass beads to the cell pellets. The cells were disrupted by shaking at 3000 rpm for 2 min with cooling using TOMMY Micro Smash MS-100R. The cell debris was removed by centrifuging at 12,000 rpm for 2 min at 4℃. 350 µl of the supernatant was mixed with an equal volume of 70 % EtOH. We transferred the mixture to the column and centrifuged it at 10,000xg for 20 seconds at 4℃. We added 350 µl of RW1 buffer to the column and centrifuged it at 10,000xg for 20 seconds at 4℃. To remove residual DNA, we added an RNase-Free DNase Set (QIAGEN, Germany) to the column. After 15 min digestion at room temperature, we added RW1 buffer to the column and centrifuged it at 10,000xg for 20 seconds at 4℃. We added 500 µl of RPE buffer to the column and centrifuged it at 10,000xg for 20 seconds at 4℃. We repeated this process twice. We collected RNA by adding 50 µl of RNase-free water to the column and centrifuging it at 10,000xg for 30 seconds at 4℃. We measured the RNA concentration by nanodrop. The quality and integrity of the RNA samples were assessed using an Agilent 2200 TapeStation (Agilent Technologies) We prepared the library using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). For indexing, we used NEBNext Multiplex Oligos for Illumina (NEB #E6440). We followed the manufacturer's protocol to prepare the libraries.
identifiers.namespace: GEO
identifiers.id: GSM8902288_r1