Patent ID: 8986958
Filing Date: 2015-03-24
Classification: C12Q

Abstract:
1. A method for generating a population of single-stranded nucleic acid probes, each probe comprising a predetermined nucleotide sequence, the method comprising: a) providing a starting population of linear double-stranded nucleic acid precursor molecules each precursor molecule having (i) a probe region having the predetermined sequence which is flanked at a 5′ and a 3′ end by a first and a second restriction enzyme recognition sequence for generating ligation substrates and for ligating a plurality of the double-stranded nucleic acid precursor molecules into head-to-tail concatemers (ii) the 5′ flanking region including the first restriction enzyme recognition sequence and (iii) the 3′ flanking region including the second restriction enzyme recognition sequence; b) contacting the 5′ and 3′ flanking regions of the linear double-stranded nucleic acid precursor molecules with a first and a second restriction enzyme to cleave the first and second restriction enzyme recognition sequences so as to generate ligation substrates; c) ligating the ligation substrates together so as to generate a plurality of random head-to-tail concatemers; d) amplifying the plurality of head-to-tail concatemers; e) contacting the amplified head-to-tail concatemers with the first and second restriction enzymes so as to release a plurality of double-stranded monomer linear precursor molecules; and f) selectively removing one strand of the double-stranded monomer linear precursor molecules so as to generate a population of single-stranded nucleic acid probes, each probe comprising the predetermined nucleotide sequence.