title: Novel tools for unbiased DNA differential methylation screening.
abstract: DNA differential methylation screening approaches may be hypothesis driven (preselection of the loci to screen) or unbiased (screening precedes mapping of differentially methylated loci). The latter allow for the identification of sequences demonstrating 'nonclassical' methylation behavior in cancer and, thus, widen our concept of tumor biology. Extensive employment of unbiased screening methods is hampered by the troublesome procedures involved in physically mapping the identified differentially methylated DNA fragments. In this special report, we describe a positive experience of optimizing two screening methods, methylation-sensitive arbitrarily primed PCR and amplification of intermethylated sites, with a special focus on simplification, or even exclusion, of sequencing procedures. With our modifications, unbiased screening of DNA differential methylation acquires an easy-to-use workflow, including sophisticated experimental design, high-resolution analysis and simple genomic mapping of the fragments of interest.
authors: Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Moskvorechie Street 1, Moscow, Russia.
authors: Alexander S
authors: AS
authors: Tanas
authors: Viktoria V
authors: VV
authors: Shkarupo
authors: Ekaterina B
authors: EB
authors: Kuznetsova
authors: Dmitry V
authors: DV
authors: Zaletayev
authors: Vladimir V
authors: VV
authors: Strelnikov
mesh_terms: D002874:Chromosome Mapping / Q000379:methods*; D019175:DNA Methylation / Q000235:genetics*; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D057166:High-Throughput Screening Assays / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004273:DNA, Neoplasm