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首页> 外国专利> Preparing genetic fingerprint, useful e.g. for identifying an individual animal, plant or microorganism, using a universal array of hybridization probes

Preparing genetic fingerprint, useful e.g. for identifying an individual animal, plant or microorganism, using a universal array of hybridization probes

机译：准备遗传指纹，例如使用通用的杂交探针阵列鉴定单个动物，植物或微生物

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Producing a genetic fingerprint of an organism, is new. Producing a genetic fingerprint of an organism comprises extracting double-stranded (ds) DNA (I), determining its concentration and, if necessary, diluting to 1-20 ng/ micro l. (I) is digested, before or after dilution, with at least one restriction enzyme (RE) having a specific restriction site (RS), to produce ds fragments (II) that include a residue from RS and an internal sequence particularly to each (II). Optionally a ds oligonucleotide adapter (ONA), of known sequence, is attached to the ends of (II), and (II) are then amplified by PCR, using a primer complementary to the residue of RS and optionally to ONA. (II) are labeled during or after amplification and then denatured to create a collection of single-stranded (ss) DNA fragments (III), optionally linked to ss-ONA. This collection is hybridized to a chip comprising, on a carrier, an array of nucleic acid probes (P), comprising two parts. P1 is the same for all probes and complementary to the residue of RS, and P2, contiguous with P1, is different for each probe and comprising a known combination of nucleotide bases so that the array can hybridize specifically to different fragments in the collection when not bound to ss-ONA. P1 is also complementary to ONA where (III) are linked to ss-ONA. The chip is washed, particularly to remove non-hybridized fragments, then dried. Independent claims are also included for the following: (1) genetic fingerprint produced by the new method; and (2) identifying an organism by comparing its fingerprint, prepared by the new method, with a reference fingerprint prepared the same way.
机译：产生生物体的基因指纹是新的。产生生物体的遗传指纹包括提取双链（ds）DNA（I），确定其浓度，并在必要时稀释至1-20 ng /μl。 （I）在稀释之前或之后用至少一种具有特定限制位点（RS）的限制酶（RE）消化，以产生ds片段（II），该片段包含RS的残基以及每个序列的内部序列（ II）。任选地，将具有已知序列的ds寡核苷酸衔接子（ONA）连接至（II）的末端，然后使用与RS残基和任选地与ONA残基互补的引物通过PCR扩增（II）。 （II）在扩增期间或之后被标记，然后变性以产生单链（ss）DNA片段（III）的集合，其任选地连接至ss-ONA。该集合与包含在载体上的包含两部分的核酸探针（P）阵列的芯片杂交。 P1对于所有探针都是相同的，并且与RS的残基互补，并且与P1相邻的P2对于每个探针都是不同的，并且包含已知的核苷酸碱基组合，因此该阵列可以与收集物中的不同片段特异性杂交绑定到ss-ONA。 P1也与ONA互补，其中（III）与ss-ONA连接。洗涤芯片，特别是去除未杂交的片段，然后干燥。还包括以下方面的独立权利要求：（1）新方法产生的遗传指纹; （2）通过比较用新方法制备的生物指纹和用相同方法制备的参考指纹来识别生物。

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