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Identifying nucleic acids by restriction digestion and hybridization with random or pseudorandom oligonucleotides
Identifying nucleic acids by restriction digestion and hybridization with random or pseudorandom oligonucleotides
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机译:通过限制性消化和与随机或伪随机寡核苷酸杂交来鉴定核酸
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摘要
This invention provides a method of determining whether a nucleic acid molecule having a sequence of interest is present within a nucleic acid- containing sample which comprises the steps: (A) subjecting said nucleic acid of said sample to a treatment sufficient to fragment said nucleic acid and to produce a plurality of fragments derived from said nucleic acid; (B) fractionating said plurality of fragments; (C) contacting said fractionated plurality of fragments, under hybridizing conditions, with a reagent comprising a mixture of random or pseudorandom oligonucleotides, said oligonucleotides having a length a ≧3 and ≦ 1000 nucleotides; (D) detecting the pattern of hybrids that form from said contacting of said fractionated nucleic acid molecules and said oligonucleotides of the mixture of random or pseudorandom oligonucleotides; and (E) determining whether said nucleic acid molecule having said sequence of interest is present in said sample by determining whether said pattern is indicative of the presence of said molecule. This invention provides a method of obtaining a signature for a nucleic acid of interest and for constructing a restriction map of a nucleic acid of interest. This invention finally provides a method of identifying a nucleic acid sequence which encodes, directly or indirectly, a particular biological activity of interest.
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