Patent ID: 6531302
Filing Date: 2003-03-11
Classification: C12Q

Abstract:
A method for the amplification of one or more target nucleic acids of interest in one or more samples using a bioelectronic microchip comprising:a) introducing at least one of the target nucleic acids onto a bioelectronic microchip having a plurality of electronically addressable capture sites; b) electronically addressing the target nucleic acids to at least one capture site which has attached thereto first and second strand displacement amplification oligonucleotide primers having the capacity to support strand displacement amplification, the first and second primers comprising a restriction endonuclease recognition sequence upstream of a sequence specific for the target nucleic acid to be amplified; c) hybridizing the target nucleic acid to be amplified to at least the first strand displacement amplification primer; d) contacting the hybridized target nucleic acid with enzymes and reagents necessary to support strand displacement amplification, including a restriction endonuclease which recognizes the restriction endonuclease recognition sequence; and e) amplifying the target nucleic acid by strand displacement amplification to produce amplicon species, wherein the amplification comprises: i) extending the first strand displacement amplification primer to which the target nucleic acid has hybridized to form a first primer amplicon comprising a nucleic acid sequence to which the target nucleic acid specific portion of the second primer hybridizes; ii) dissociating the first primer amplicon from target nucleic acid in at least the region to which the target nucleic acid specific portion of the second primer hybridizes; and iii) hybridizing at least a portion of the first primer amplicons to proximate second strand displacement amplification primers and extending the proximate second strand displacement amplification primers to form second primer amplicons which are hybridized to the endonuclease recognition site of the first primer amplicon, thereby producing an amplicon capable of supporting strand-displacement self-amplification.