biostudies-literatureUnknown13Baumann VWe aimed at extending the repertoire of high-quality miRNA normalizers for reverse transcription-quantitative PCR (RT-qPCR) of human plasma with special emphasis on the extremely guanine-cytosine-rich portion of the miRNome. For high-throughput selection of stable candidates, microarray technology was preferred over small-RNA sequencing (sRNA-seq) since the latter underrepresented miRNAs with a guanine-cytosine (GC) content of at least 75% (<i>p</i> = 0.0002, <i>n</i> = 2). miRNA abundances measured on the microarray were ranked for consistency and uniformity using nine normalization approaches. The eleven most stable sequences included miRNAs of moderate, but also extreme GC content (45%-65%: miR-320d, miR-425-5p, miR-185-5p, miR-486-5p; 80%-95%: miR-1915-3p, miR-3656-5p, miR-3665-5p, miRFrontiers in genetics1058668https://www.ebi.ac.uk/biostudies/studies/S-EPMC9846067biostudies-literatureIdentification of extremely GC-rich micro RNAs for RT-qPCR data normalization in human plasma.PMC9846067Schwerdtfeger ARSteinborn RBaumann VFaridani ORAthanasiou ATWallner BfalseIdentification of extremely GC-rich micro RNAs for RT-qPCR data normalization in human plasma.We aimed at extending the repertoire of high-quality miRNA normalizers for reverse transcription-quantitative PCR (RT-qPCR) of human plasma with special emphasis on the extremely guanine-cytosine-rich portion of the miRNome. For high-throughput selection of stable candidates, microarray technology was preferred over small-RNA sequencing (sRNA-seq) since the latter underrepresented miRNAs with a guanine-cytosine (GC) content of at least 75% (<i>p</i> = 0.0002, <i>n</i> = 2). miRNA abundances measured on the microarray were ranked for consistency and uniformity using nine normalization approaches. The eleven most stable sequences included miRNAs of moderate, but also extreme GC content (45%-65%: miR-320d, miR-425-5p, miR-185-5p, miR-486-5p; 80%-95%: miR-1915-3p, miR-3656-5p, miR-3665-5p, miR2022-01-01T00:00:00Z20222025-04-21T16:42:22.203Z2025-02-19T02:18:02.244ZS-EPMC98460673668585410.3389/fgene.2022.1058668