Determining DNA polymorphisms and cytosine methylation, comprises a two-step PCR amplification and hybridization to an array of oligonucleotides and/or peptide nucleic acids and chemical and/or enzymatic treatment

摘要

Determining DNA polymorphisms and cytosine methylation in a DNA molecule (I), comprising: (a) chemical and/or enzymatic treatment of (I) for cytosine methylation analysis; (b) PCR amplification of (I); (c) hybridization of amplicons to immobilized oligonucleotides and/or peptide nucleic acid (PNA) oligomers; and (d) from the hybridization patterns, drawing conclusions about the sequence features and methylation patterns of (I), is new. Determining DNA polymorphisms and cytosine methylation of a DNA molecule (I), comprising: (a) separating (I) from a sample; (b) for cytosine methylation analysis, chemical and/or enzymatic treatment of (I); (c) amplification of (I) in a polymerase reaction, using one primer in solution and a second primer bound to a solid phase, where at least one primer comprises two domains, where the 3'-domain can hybridize to (I) while the 5'-domain can not; (d) amplifying the amplified (I) again using primers that hybridize to the 5'-domain of at least one primer of step (c), or identical with it, and are detectably labeled; and (e) hybridizing amplicons to immobilized oligonucleotides and/or PNA (peptide nucleic acid) oligomers, unable to function as primers; and (f) from the hybridization pattern, drawing conclusions about the sequence features and methylation patterns of (I), is new. An Independent claim is also included for a device, comprising: (a) a surface on which are immobilized primer oligonucleotides (attached through the 5'-end) and other oligonucleotides or PNA oligomers that can not be extended in a polymerase reaction; (b) a chamber in which the surface of (a) forms one wall; (c) a system for controlling the temperature of the chamber; and (d) a system for supplying liquid to the chamber.