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Identifying nucleic acids by restriction digestion and hybridization with random or pseudorandom oligonucleotides

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

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.
机译:本发明提供了一种确定含有核酸的样品中是否存在具有目的序列的核酸分子的方法,该方法包括以下步骤:(A)对所述样品的所述核酸进行足以使所述核酸片段化的处理。并产生多个衍生自所述核酸的片段; (B)分级分离所述多个片段; (C)在杂交条件下,使所述分级分离的多个片段与包含随机或伪随机寡核苷酸的混合物的试剂接触,所述寡核苷酸的长度为≧ 3和≦ 1000个核苷酸; (D)检测由所述分级核酸分子与所述随机或伪随机寡核苷酸的混合物的所述寡核苷酸的所述接触形成的杂交体的模式; (E)通过确定所述模式是否表示所述分子的存在来确定所述样品中是否存在具有所述目的序列的核酸分子。本发明提供了一种获得目的核酸的标记并构建目的核酸的限制性图谱的方法。本发明最终提供了鉴定直接或间接编码目的特定生物学活性的核酸序列的方法。

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