Figure 4 | Scientific Reports

Figure 4

From: Targeted sequencing of both DNA strands barcoded and captured individually by RNA probes to identify genome-wide ultra-rare mutations

Figure 4

Performance evaluation of DEEPER-Capture. (A) For each of the six DEEPER-Libraries derived from different amounts (500 ng, 20 ng, 1 ng, 100 pg, 20 pg and 10 pg) of input genomic DNA, enrichment efficiencies of 298 cancer related genes were calculated. Recovery ratios of DEEPER-Capture and Half-DEEPER-Capture for all 298 genes in six libraries were quantified by real-time PCR assays detecting each gene’s abundance in the libraries before and after DEEPER-Capture or Half-DEEPER-Capture. (B) An insert sequence composed of amplicon regions of five genes whose GC contents fall into a broad range (27.3% to 74.1%) was cloned into a pcDNA vector. (C) Real-time PCR analysis of sequential dilutions of the plasmid. 1, 10, 100, 1,000 and 10,000 femtomoles of the plasmids were added as templates for the assays. C t value for each gene observed from different plasmid template amount was plotted, and trend lines were shown. No significant GC-dependent amplification bias was observed for real-time PCR assays. (D) A whole genome DEEPER-Library, and whole exome DEEPER-Libraries captured by Half-DEEPER-Capture and DEEPER-Capture were analyzed on an agarose gel.

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