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2. 发明公开

EP1186671A2 Method for hybridization of arrays on siliceous surfaces 审中-公开
标题翻译： Methode zur Hybridisierung von Arrays aufSilikonoberflächen
公开(公告)号：EP1186671A2
公开(公告)日：2002-03-13
申请号：EP01121185.1
申请日：2001-09-04
申请人： Agilent Technologies Inc. (a Delaware Corporation)
发明人： Shannon, Karen W. , Lefkowitz, Steven M.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6837 , C12Q2527/119 , C12Q2527/101
摘要： A method (100, 200) of hybridizing arrays of nucleic acids an surface-derivatized siliceous substrates with other nucleic acid materials provides an envelope of conditions to produce sensitive, selective detection of nucleic acid targets, while preserving the intactness of the derivatized surface of the array. The envelope of hybridization conditions includes a hybridization solution having a pH between pH 5.5 and 6.7 and a high hybridization or incubation temperature between 55°C and 70°C. In one embodiment (100), the hybridization solution is maintained (102) at the pH between pH 5.5 and 6.7 and the array is incubated (104) with a nucleic acid material in the pH-maintained hybridization solution at the hybridization temperature of between 55°C and 70°C. The pH of the hybridization solution is maintained (102) with a buffer having a useful buffering capacity between pH 5.5 and 6.7. In another embodiment (200), a nucleic acid material is combined (202) with the hybridization solution at the pH between pH 5.5 to 6.7 containing a buffer and a monovalent cation and the combined solution is incubated (204) with the array at the hybridization temperature of between 55°C and 70°C so as to hybridize the nucleic acid material. Typical hybridization times can range from less than 2 hours to more than 48 hours. The present method is particularly useful for hybridization assays an silylated-siliceous substrates where the incubation time is much greater than about 6 hours at the high hybridization temperature. The envelope of hybridization conditions provide optimized assay performance while maintaining the integrity of the derivatized surface of the siliceous substrate. A kit comprises a microarray having a siliceous substrate with a derivatized surface and an oligonucleotide populated an the surface for hybridizing a another oligonucleotide material. The kit further includes instructions for using the microarray in accordance with the method (100, 200) of the present invention.
摘要翻译：将表面衍生的硅质底物与其他核酸材料的核酸阵列杂交的方法（100,200）提供条件的包络，以产生对核酸靶的敏感，选择性检测，同时保持衍生化表面的完整性阵列。杂交条件的包络包括pH在5.5和6.7之间的pH值和55℃至70℃之间的高杂交或孵育温度的杂交溶液。在一个实施方案（100）中，将杂交溶液维持在（102）在pH 5.5和6.7之间的pH值，并且在55℃至70℃的杂交温度下，将该阵列与含有pH值的杂交溶液中的核酸材料一起孵育（104）。保持杂交溶液的pH（102 ）与具有在5.5和6.7之间的有用缓冲能力的缓冲液。在另一个实施方案（200）中，将核酸材料与杂交溶液在含有缓冲液和一价阳离子的pH5.5至6.7之间的pH下结合（202），并将合并的溶液与杂交（204）与阵列孵育温度在55℃至70℃之间，以使核酸材料杂交。典型的杂交时间可以从小于2小时到超过48小时。本方法对于硅烷化硅质底物的杂交测定特别有用，其中在高杂交温度下孵育时间远大于约6小时。杂交条件的包络提供优化的测定性能，同时保持硅质底物的衍生化表面的完整性。试剂盒包括具有带有衍生化表面的硅质底物的微阵列和用于杂交另一种寡核苷酸材料的表面的寡核苷酸。该试剂盒还包括根据本发明的方法（100,200）使用微阵列的说明书。

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