1. Schematic of experimental parameters comparing FFPE DNA repair methods to FF DNA. DNA (500 ng) from FF tissue was processed according to Illumina Human Methylation instructions, including bisulfite modification followed by the standard Human Methylation pro...
Schematic of experimental parameters comparing FFPE DNA repair methods to FF DNA. DNA (500 ng) from FF tissue was processed according to Illumina Human Methylation instructions, including bisulfite modification followed by the standard Human Methylation processing protocol (Gold standard). Experimental conditions are separated for REPLI-g ligation (LIG) and Illumina Restore Kit (RES). LIG1: the original Thirwell method using 500 ng of genomic DNA processed by REPLI-g ligase and bisulfite modified (BS), and 4 μL of bisulfite-modified DNA used for the starting material for the Illumina Human Methylation array kit. LIG2: Thirwell method with output DNA increased to 8 μL of bisulfite-modified DNA, which is the same as used in the Restore Kit. LIG3 and LIG4: 500 ng and 250 ng of genomic DNA, respectively, were bisulfite modified, processed by REPLI-g ligase and 8 μL of material used for Illumina Methylation Array kit. RES1: the Illumina Restore protocol using 500 ng of genomic DNA, bisulfite modified, and processed per Restore Kit protocol (including 8 μL for Array steps). RES2 and RES3: technical replicates for Restore Kit using 250 ng of genomic DNA.
Erin M. Siegel et al. Cancer Epidemiol Biomarkers Prev 2014;23:
©2014 by American Association for Cancer Research