GSM3503272
GSM3503272: EM_low_female_R2; Danio rerio; RNA-Seq
Total RNA were extracted using Trizol method. We added 1 ml Trizol (invitrogen) to the sample (4-7 million cells) stored at -80 °C and homogenized the sample by pipette. Then, 200 ul Chloroform was added to the homogenate and the sample was vigorously shaked by hand for 15 seconds until the phase separation disappeared. After standing at room temperature for 2-3 minutes, the sample was centrifuged at 12,000 g for 15 minutes at 4°C. The colorless supernatant was transferred to another new centrifuge tube, mixed with equal volume of isopropanol thoroughly, kept standing at -20°C for 10 minutes, and then centrifuged at 12,000 g for 10 minutes at 4°C to pellet the RNA. The supernatant was carefully removed, and the pellet was gently washed with 1 ml fresh 75% ethanol for 2-3 times, dried for 2 to 3 minutes at room temperature and dissolved in 50 ul RNase-free water. Stranded PolyA+ RNA libraries were prepared at BGI with in-house kits. The quality of the libraries were assessed by checking the distribution of the fragment size using the Agilent 2100 bioanalyzer, and the quantity of the libraries were measured using real-time quantitative PCR (TaqMan Probe). The qualified libraries were amplified on cBot to generate clusters on the flowcell, and then sequenced on the Illumina HiSeq XTen System.
GEO
GSM3503272
GEO Accession
GSM3503272