Figure 3
From: Methylation-capture and Next-Generation Sequencing of free circulating DNA from human plasma
Size distribution of fcDNA in 5 control samples and following fcDNA library construction (A and B) and DNA recovery following MBD-capture (C). Agilent Bioanalyzer chip with fcDNA isolated from 5 control subjects prior to (A) and following (B) methylation enrichment and library preparation. Increase in molecular weight reflects successful adapter ligation. MWM: molecular weight marker. DNA recovery following MBD-capture (C). Recovery of DNA (based on % of input DNA) as quantitated by qPCR following the standard (grey bars) or our modified (white bars) MBD-capture protocol. SFTA3 promoter is unmethylated in PBMC DNA and methylated in the methylation positive control SSSI. At low DNA inputs (100 ng), SFTA3 is recovered from both PBMC and SSSI DNA, suggesting high background MBD binding and low methylation enrichment. With our modified protocol, SFTA3 is minimally recovered from PBMC DNA (unmethylated) but is recovered from SSSI DNA (methylated), even at low DNA inputs. Data is presented as % DNA recovery ± range.