Figure 1
From: MitoRCA-seq reveals unbalanced cytocine to thymine transition in Polg mutant mice
An overview of the mitoRCA-seq procedure.
(A) Mitochondrial DNA is enriched by RCA using mtDNA specific primers in a single tube. Further enrichment of mtDNA fragments is achieved by restriction enzyme digestion (EcoRV for mouse) and only full-length mouse mtDNA can give rise to two discrete DNA fragments (mouse: 9.5 kb and 6.8 kb). Different starting materials (1 ng, 5 ng and 50 ng total DNA, from left to right) were tested. (B) The computational workflow employed to identify mtDNA mutations. For raw reads with the same sequence, only one read was kept for downstream mutation calling, which has the highest average quality score (defined as best-unique read). (C) The overall correlation of three technical replicates derived from the liver of a Polg mutant mouse. The frequencies of individual mutations identified in each library are plotted. RCA1-1 and RCA1-2 represents two libraries prepared from the same RCA reaction. RCA2 is a technical replicate prepared from a different RCA reaction. The correlation coefficient (R) between technical replicates is 0.998 (RCA1-1 and RCA1-2), 0.996 (RCA1-1 and RCA2) and 0.997 (RCA1-2 and RCA2), respectively.
