GSM4512167
GSM4512167: Senescent HUVECs + Metformin replicate 3; Homo sapiens; miRNA-Seq
Total RNA, including small (< 200 nucleotides) RNAs, was extracted from HUVEC pellets using Norgen's total RNA Purification Kit (cat. no. 37500, Norgen Biotek Corporation, Canada) according to the manufacturer's protocol. Purified RNA was stored at −80°C until analysis. Small RNA sequencing was performed in triplicate on Young and SEN cells, and SEN cells treated with metformin. TruSeq Small RNA Library PrepKit v2 (Illumina; RS-200-0012/24/36/48) was used for library preparation according to the manufacturer's indications. Briefly, 35 ng purified RNA was linked to RNA 3' and 5' adapters, converted to cDNA, and amplified using Illumina primers containing unique indexes for each sample. Each library was quantified using Agilent Bioanalyzer and High Sensitivity DNA Kit (cat. no. 5067-4626, Agilent Technologies, USA) and equal amounts of libraries were pooled together. Size selection allowed keeping 130-160 bp fragments. After ethanol precipitation, the library pool was quantified with Agilent High Sensitivity DNA Kit, diluted to 1.8 pM, and sequenced using NextSeq® 500/550 High Output Kit v2 (75 cycles) (Illumina; FC-404-2005) on the Illumina NextSeq500 platform.
GEO
GSM4512167
GEO Accession
GSM4512167