GSM6248699: blood, P-3; Homo sapiens; RNA-Seq
Samples were collected using EDTA anticoagulated blood collection vessels. Take a 15 ml tube, add 6 ml trizol and 2 ml fresh blood (trizol: blood =3:1), and blow several times with a pipette to help split the cells in the sample. The sample oscillates violently and mixes 1-2 min until all the flocs were dissolved.Incubate at room temperature for 5 min to completely decompose the ribosomes. The samples were stored in the refrigerator at -80℃ for long-term storage for high-throughput sequencing. A total amount of 3 μg RNA per sample was used as input material for the RNA sample preparations. Firstly,ribosomal RNA was removed by Epicentre Ribo-zero™ rRNA Removal Kit (Epicentre, USA), and rRNA free residue was cleaned up by ethanol precipitation. Subsequently, sequencing libraries were generated using the rRNA-depleted RNA by NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations.