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公开(公告)号：US20090246760A1
公开(公告)日：2009-10-01
申请号：US12058032
申请日：2008-03-28
CPC分类号： C12Q1/6851 , C12Q2537/165
摘要： The disclosure provides methods of reducing the range of representation levels of nucleic acid targets. The methods are particularly useful for multi-target analyses benefiting from a low variance of target representations, such as, e.g., single molecule sequencing and/or heterozygous genotyping, and pathogen diagnosis. Two general methods are provided. In Method 1, starting concentrations of probes are adjusted. In Method 2, target-specific probes are “binned,” i.e., several subsets of probes are selected based on similar representation levels. Thereafter, each subset of corresponding targets is extracted, with or without amplification, using a separate portion of the sample (i.e., separate vessels).
摘要翻译： 本公开提供了减少核酸靶标的表达水平范围的方法。 这些方法对于受益于靶表现的低方差（例如单分子测序和/或杂合基因分型）和病原体诊断的多靶标分析特别有用。 提供了两种一般方法。 在方法1中，调节探针的起始浓度。 在方法2中，目标特异性探针被“分类”，即，基于相似的表示级别来选择几个探针子集。 此后，使用样本的单独部分（即，分离的血管），提取具有或不具有扩增的相应靶标的每个子集。
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公开(公告)号：US20090156412A1
公开(公告)日：2009-06-18
申请号：US11958173
申请日：2007-12-17
CPC分类号： C12N15/1006 , C12Q1/6806 , C12Q1/6874 , C12Q2565/537 , C12Q2525/307 , C12Q2521/319
摘要： The disclosure provides methods of capturing target nucleic acids (e.g., gene or gene fragments) onto a solid support for further analysis. The disclosed methods utilize a capture probe that selectively circularizes only the target nucleic acid. Following the circularization of the target, the linear, non-target, nucleic acids are removed from the sample. Next, the circularized target is linearized and bound to a solid support. To allow for linearization, the capture probe may include a cleavage site that can be a noncanonical nucleotide(s) (e.g., uracil in DNA) and/or a rare-cutter site (e.g., the Not I restriction site). In some embodiments, the target nucleic acid is captured onto a support without an intermediate amplification step.
摘要翻译： 本公开提供了将靶核酸（例如，基因或基因片段）捕获到固体支持物上用于进一步分析的方法。 所公开的方法利用仅选择性地环化靶核酸的捕获探针。 在靶标的圆化之后，从样品中除去线性非靶标核酸。 接下来，环化的靶被线性化并结合到固体支持物上。 为了允许线性化，捕获探针可以包括可以是非科学核苷酸（例如DNA中的尿嘧啶）和/或稀有切割位点（例如Not I限制性位点）的切割位点。 在一些实施方案中，将目标核酸捕获到载体上而无需中间扩增步骤。
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公开(公告)号：US20110027790A1
公开(公告)日：2011-02-03
申请号：US12871978
申请日：2010-08-31
CPC分类号： C12Q1/6851 , C12Q2537/165
摘要： The disclosure provides methods of reducing the range of representation levels of nucleic acid targets. The methods are particularly useful for multi-target analyses benefiting from a low variance of target representations, such as, e.g., single molecule sequencing and/or heterozygous genotyping, and pathogen diagnosis. Two general methods are provided. In Method 1, starting concentrations of probes are adjusted. In Method 2, target-specific probes are “binned,” i.e., several subsets of probes are selected based on similar representation levels. Thereafter, each subset of corresponding targets is extracted, with or without amplification, using a separate portion of the sample (i.e., separate vessels).
摘要翻译： 本公开提供了减少核酸靶标的表达水平范围的方法。 这些方法对于受益于靶表现的低方差（例如单分子测序和/或杂合基因分型）和病原体诊断的多靶标分析特别有用。 提供了两种一般方法。 在方法1中，调节探针的起始浓度。 在方法2中，目标特异性探针被“分类”，即，基于相似的表示级别来选择几个探针子集。 此后，使用样本的单独部分（即，分离的血管），提取具有或不具有扩增的相应靶标的每个子集。
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公开(公告)号：US07790391B2
公开(公告)日：2010-09-07
申请号：US12058032
申请日：2008-03-28
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6851 , C12Q2537/165
摘要： The disclosure provides methods of reducing the range of representation levels of nucleic acid targets. The methods are particularly useful for multi-target analyses benefiting from a low variance of target representations, such as, e.g., single molecule sequencing and/or heterozygous genotyping, and pathogen diagnosis. Two general methods are provided. In Method 1, starting concentrations of probes are adjusted. In Method 2, target-specific probes are “binned,” i.e., several subsets of probes are selected based on similar representation levels. Thereafter, each subset of corresponding targets is extracted, with or without amplification, using a separate portion of the sample (i.e., separate vessels).
摘要翻译： 本公开提供了减少核酸靶标的表达水平范围的方法。 这些方法对于受益于靶表现的低方差（例如单分子测序和/或杂合基因分型）和病原体诊断的多靶标分析特别有用。 提供了两种一般方法。 在方法1中，调节探针的起始浓度。 在方法2中，目标特异性探针被“分类”，即，基于相似的表示级别来选择几个探针子集。 此后，使用样本的单独部分（即，分离的血管），提取具有或不具有扩增的相应靶标的每个子集。
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