title: Single DNA molecule patterning for high-throughput epigenetic mapping.
abstract: We present a method for profiling the 5-methyl cytosine distribution on single DNA molecules. Our method combines soft-lithography and molecular elongation to form ordered arrays estimated to contain more than 250 000 individual DNA molecules immobilized on a solid substrate. The methylation state of the DNA is detected and mapped by binding of fluorescently labeled methyl-CpG binding domain peptides to the elongated dsDNA molecules and imaging of their distribution. The stretched molecules are fixed in their extended configuration by adsorption onto the substrate so analysis can be performed with high spatial resolution and signal averaging. We further prove this technique allows imaging of DNA molecules with different methylation states.
authors: School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, United States.
authors: Aline
authors: A
authors: Cerf
authors: Benjamin R
authors: BR
authors: Cipriany
authors: Jaime J
authors: JJ
authors: Benítez
authors: Harold G
authors: HG
authors: Craighead
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics*; D018899:CpG Islands; D003596:Cytosine / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D018983:DNA Footprinting; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism; D057890:Epigenomics; D057166:High-Throughput Screening Assays; D007091:Image Processing, Computer-Assisted; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D004268:DNA-Binding Proteins; D003596:Cytosine; D004247:DNA