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    • 7. 发明申请
    • MOSTLY NATURAL DNA SEQUENCING BY SYNTHESIS
    • 通过合成测序的大多数天然DNA
    • WO2010117804A3
    • 2011-06-09
    • PCT/US2010029238
    • 2010-03-30
    • UNIV CALIFORNIAHUANG XIAOHUAROLLER ERIC
    • HUANG XIAOHUAROLLER ERIC
    • C12Q1/68C12N15/11G01N33/52
    • C12Q1/6869C12Q2535/119C12Q2525/301C12Q2527/137C12Q2535/113
    • The invention provides a new method for DNA sequencing called "natural sequencing by synthesis" (nSBS). According to the method, DNA that includes a desired sequence is synthesized using a dNTP mix with a small percentage of fluorescently-labeled nucleotides. The fluorescent label is cleavable. In contrast to previous methods that utilize 100% labeled nucleic acids, use of a small percentage of labeled nucleic acids minimizes the distortion of the natural structure of the extending DNA strand and the DNA polymerase. Using the disclosed methods with less than 10,000 copies of template DNA and 10% of the nucleotides labeled, long homopolymer stretches up to 20 bases can be sequenced with high accuracy and Q20 (with 99% accuracy) read lengths of up to 1,000 bases can be achieved. A Q20 read length of greater than 100 bases can potentially be achieved, even if the sequencing is performed with 1,000 copies of a template and 10% of the nucleotides labeled.
    • 本发明提供了一种称为“合成天然测序”(nSBS)的DNA测序的新方法。 根据该方法,使用具有小百分比的荧光标记核苷酸的dNTP混合物合成包含所需序列的DNA。 荧光标记是可切割的。 与使用100%标记的核酸的先前方法相比,使用小百分比的标记的核酸使延伸的DNA链和DNA聚合酶的天然结构的变形最小化。 使用所公开的方法,使用少于10,000拷贝的模板DNA和10%标记的核苷酸,长均聚物延伸多达20个碱基可以高精度测序,Q20(具有99%的准确度)读取长度可达1,000个碱基可以 实现。 即使用1000份拷贝的模板和10%的标记的核苷酸进行测序,潜在地可以实现大于100个碱基的Q20读取长度。
    • 8. 发明申请
    • MOSTLY NATURAL DNA SEQUENCING BY SYNTHESIS
    • 通过合成测序的大多数天然DNA
    • WO2010117804A2
    • 2010-10-14
    • PCT/US2010/029238
    • 2010-03-30
    • THE REGENTS OF THE UNIVERSITY OF CALIFORNIAHUANG, XiaohuaROLLER, Eric
    • HUANG, XiaohuaROLLER, Eric
    • C12Q1/68C12N15/11G01N33/52
    • C12Q1/6869C12Q2535/119C12Q2525/301C12Q2527/137C12Q2535/113
    • The invention provides a new method for DNA sequencing called "natural sequencing by synthesis" (nSBS). According to the method, DNA that includes a desired sequence is synthesized using a dNTP mix with a small percentage of fluorescently-labeled nucleotides. The fluorescent label is cleavable. In contrast to previous methods that utilize 100% labeled nucleic acids, use of a small percentage of labeled nucleic acids minimizes the distortion of the natural structure of the extending DNA strand and the DNA polymerase. Using the disclosed methods with less than 10,000 copies of template DNA and 10% of the nucleotides labeled, long homopolymer stretches up to 20 bases can be sequenced with high accuracy and Q20 (with 99% accuracy) read lengths of up to 1,000 bases can be achieved. A Q20 read length of greater than 100 bases can potentially be achieved, even if the sequencing is performed with 1,000 copies of a template and 10% of the nucleotides labeled.
    • 本发明提供了一种称为“合成天然测序”(nSBS)的DNA测序的新方法。 根据该方法,使用具有小百分比的荧光标记核苷酸的dNTP混合物合成包含所需序列的DNA。 荧光标记是可切割的。 与使用100%标记的核酸的先前方法相比,使用小百分比的标记的核酸使延伸的DNA链和DNA聚合酶的天然结构的变形最小化。 使用所公开的方法,使用少于10,000拷贝的模板DNA和10%标记的核苷酸,长均聚物延伸多达20个碱基可以高精度测序,Q20(具有99%的准确度)读取长度可达1,000个碱基可以 实现。 即使用1000份拷贝的模板和10%的标记的核苷酸进行测序,潜在地可以实现大于100个碱基的Q20读取长度。