title: GSM5369459: ECA-11; Homo sapiens; miRNA-Seq
library_construction_protocol: According to the manufacturer's instructions, TRIzol (Invitrogen, CA, USA) was used to extract total RNA from frozen tissue. Use NanoDrop ND-1000 instrument (NanoDrop-Thermo, DE, USA) to evaluate the quantity and concentration of each RNA sample, and check the integrity by agarose gel electrophoresis. Before preparing the total RNA sample library, perform the following treatments to remove the modification of RNA interference small RNA sequence library construction: 3-aminoacyl (charged) deacylation to 3-OH for 3-adapter connection; 3-cP (2,3-cyclic phosphoric acid) was removed to 3-OH for 3-adapter connection; 5-OH (hydroxyl group) was phosphorylated to 5-P for 5-Adapter connection; m1A and m3C are removed for effective reverse transcription. The size of the sequencing library is selected for the RNA biotype, and the sequencing gel cutter was performed with an automatic sequencer, including 3 adaptor and 5'- adaptor connection, cDNA synthesis and library PCR amplification. The quality of the library was checked using Agilent Bioanalyzer 2100 (Invitrogen, CA, USA).