GSM6046604: Low Se1; Medicago sativa; ncRNA-Seq
GSM6046604
Complete RNA was collected using TRK-1001. A Bioanalyzer 2100 and the RNA 6000 Nano LabChip Kit (Agilent Technologies, Santa Clara, CA, United States) with RIN number > 7.0. The NanoDrop ND-1000 (NanoDrop, Wilmington, DE, USA) was used to test the volume and purity of RNA. Approximately 5 μg of total RNA was used to deplete ribosomal RNA according to the manuscript of the Ribo-Zero™ rRNA Removal Kit (Illumina, San Diego, USA). After removing ribosomal RNAs, the left RNAs were fragmented into small pieces using divalent cations under high temperatures.  Then the cleaved RNA fragments were reverse-transcribed to create the cDNA, which was next used to synthesize U-labeled second-stranded DNAs with E. coli DNA polymerase I, RNase H, and dUTP.