Validation of DNA methylation profiling in formalin-fixed paraffin-embedded samples using the Infinium HumanMethylation450 Microarray

A formalin-fixed paraffin-embedded (FFPE) sample usually yields highly degraded DNA, which limits the use of techniques requiring high-quality DNA, such as Infinium Methylation microarrays. To overcome this restriction, we have applied an FFPE restoration procedure consisting of DNA repair and ligation processes in a set of paired fresh-frozen (FF) and FFPE samples. We validated the FFPE results in comparison with matched FF samples, enabling us to use FFPE samples on the Infinium HumanMethylation450 Methylation array.     

The rfaD gene codes for ADP-L-glycero-D-mannoheptose-6-epimerase. An enzyme required for lipopolysaccharide core biosynthesis

The rfaD gene product, ADP-L-glycero-D-mannoheptose-6-epimerase, is necessary for the conversion of ADP-D-glycero-D-mannoheptose to ADP-L-glycero-D-mannoheptose. The nucleotide ADP-D-glycero-D-mannoheptose accumulates in rfaD mutant strains. Two chimeric colicin E1 plasmids carrying the coding sequence of the rfaD+ locus have been selected and shown to complement the rfaD phenotype. These clones also result in an amplification of ADP-L-glycero-D-mannoheptose-6-epimerase activity.