GSM9194195: reflected amnion protein fraction (AM173) biol rep 3; Homo sapiens; ncRNA-Seq
Total RNA was extracted from 200 µL EV or protein fraction, or tissue, using the miRNeasy Mini Kit. RNA was eluted in 30 µL nuclease-free water and stored at -80 °C until further analysis. For both the EV and protein fraction, 8.5 µL total RNA were used as input for the generation of small RNA sequencing libraries, while for the tissue, 100 ng total RNA were used. Libraries were generated with the RealSeq Biofluids library preparation kit (RealSeq Biosciences). To each sample, 1 µL miND spike-in standards were added during the first step. Adapter-ligated libraries were amplified (20 cycles for EV & protein fractions; 19 cycles for tissue) using barcoded Illumina reverse primers in combination with the Illumina forward primer. Sequencing was performed on an Illumina NovaSeq SP SR100 (Illumina, USA).