GSM8787991: Lemna aequinoctialis, control, 1; Lemna aequinoctialis; RNA-Seq
The plant RNA extraction was performed using the NucleoSpin RNA Plant Mini Kit. A powdered plant sample (100 mg) was mixed with 500 μL of Lysis buffer and 5 μL of β-mercaptoethanol, vortexed for 15 seconds, and incubated at room temperature for 3 minutes. After centrifugation at 13,000 rpm for 1 minute, 300 μL of the supernatant was transferred to a new tube, mixed with 300 μL of Membrane Lysis buffer, and filtered using an EzPure filter, followed by centrifugation at 13,000 rpm for 1 minute. Absolute ethanol (250 μL) was added to the filtrate, vortexed, and loaded onto a mini-column, which was centrifuged at 13,000 rpm for 1 minute. The column was washed with 500 μL of Wash buffer, and DNase I (70 μL) was applied for 10 minutes at room temperature for gDNA digestion. After centrifugation at maximum speed, the RNA was eluted with 50 μL of nuclease-free water and stored at –70°C. RNA sequencing libraries were constructed using the TruSeq Stranded mRNA Library Prep Kit for 150-bp paired-end sequencing on the Illumina platform.