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Fig. 2 | BMC Genomics

Fig. 2

From: Global accumulation of circRNAs during aging in Caenorhabditis elegans

Fig. 2

Experimental validation of circRNAs. a RT-PCR strategy to detect circRNAs exclusively using outward facing primer sets. Sanger sequencing of PCR products confirmed 10/10 circRNAs tested (Additional file 4: Table S2). b RT-qPCR experiments on RNase R treated mixed age adult worms. Equal amounts of mock-treated and RNase R treated RNA were used for cDNA preparation prior to qPCR. Note the enrichment of circRNAs with RNase R treatment, whereas linear cdc-42 mRNA is not enriched. c RNA-seq track visualized using Integrated Genomics Viewer from D-7 worms showing read pileup at the crh-1 gene. Note the increased read number overlapping the circularized exon. cel_circ_0000438 and cel_circ_0000439 differ by 6 nucleotides in length at the 5′ end of the exon. d Northern blot using a probe overlapping the circularized exon of crh-1 (see panel c) detects bands corresponding to circRNA and mRNA from mixed age adult RNA. Relative circRNA to mRNA abundance is enriched in RNase R treated and polyA- samples compared to polyA+ samples. e Northern blot performed using a probe that detects afd-1 circRNA and mRNA. Red arrows denote circRNA bands. Black arrows denote likely linear RNA bands

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