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    • 6. 发明申请
    • INTEGRATED SENSOR ARRAYS FOR BIOLOGICAL AND CHEMICAL ANALYSIS
    • 用于生物和化学分析的集成传感器阵列
    • WO2010047804A1
    • 2010-04-29
    • PCT/US2009005745
    • 2009-10-22
    • ION TORRENT SYSTEMS INCROTHBERG JONATHAN MSCHULTZ JONATHAN CJOHNSON KIM LREARICK TODD MMILGREM MARK JAMESMARRAN DAVIDBUSTILLO JAMES M
    • ROTHBERG JONATHAN MSCHULTZ JONATHAN CJOHNSON KIM LREARICK TODD MMILGREM MARK JAMESMARRAN DAVIDBUSTILLO JAMES M
    • G01N27/416C12M1/34
    • G01N27/4145
    • The invention is directed to apparatus and chips comprising a large scale chemical field effect transistor arrays that include an array of sample-retaining regions capable of retaining a chemical or biological sample from a sample fluid for analysis. In one aspect such transistor arrays have a pitch of 10 µm or less and each sample-retaining region is positioned on at least one chemical field effect transistor which is configured to generate at least one output signal related to a characteristic of a chemical or biological sample in such sample-retaining region. In one embodiment, the characteristic of said chemical or biological sample is a concentration of a charged species and wherein each of said chemical field effect transistors is an ion-sensitive field effect transistor having a floating gate with a dielectric layer on a surface thereof, the dielectric layer contacting said sample fluid and being capable of accumulating charge in proportion to a concentration of the charged species in said sample fluid. In one embodiment such charged species is a hydrogen ion such that the sensors measure changes in pH of the sample fluid in or adjacent to the sample-retaining region thereof. Apparatus and chips of the invention may be adapted for large scale pH-based DNA sequencing and other bioscience and biomedical applications.
    • 本发明涉及包括大规模化学场效应晶体管阵列的装置和芯片,其包括能够保留来自样品流体的化学或生物样品用于分析的样品保持区阵列。 在一个方面,这种晶体管阵列具有10μm或更小的间距,并且每个采样保持区域位于至少一个化学场效应晶体管上,该晶体管被配置为产生与化学或生物样品的特性相关的至少一个输出信号 在这样的样品保留区域中。 在一个实施方案中,所述化学或生物样品的特征是带电物质的浓度,并且其中每个所述化学场效应晶体管是离子敏感场效应晶体管,其具有在其表面上具有介电层的浮置栅极, 电介质层与所述样品流体接触并且能够与所述样品流体中的带电物质的浓度成比例地积累电荷。 在一个实施方案中,这种带电物质是氢离子,使得传感器测量样品流体在其样品保持区域中或其附近的pH值的变化。 本发明的装置和芯片可适用于大规模的基于pH的DNA测序和其他生物科学和生物医学应用。
    • 10. 发明申请
    • DETECTION AND CONFIRMATION OF NUCLEIC ACID SEQUENCES BY USE OF OLIGONUCLEOTIDES COMPRISING A SUBSEQUENCE HYBRIDIZING EXACTLY TO A KNOWN TERMINAL SEQUENCE AND A SUBSEQUENCE HYBRIDIZING TO AN UNIDENTIFIED SEQUENCE
    • 核苷酸序列的检测和确认通过使用包含随机杂交的寡核苷酸完全杂交到已知的终末序列和随机杂交到未经鉴别的序列
    • WO9907896A3
    • 1999-04-29
    • PCT/US9816548
    • 1998-08-07
    • CURAGEN CORPROTHBERG JONATHAN MDEEM MICHAEL WSIMPSON JOHN W
    • ROTHBERG JONATHAN MDEEM MICHAEL WSIMPSON JOHN W
    • C12N15/09C12N15/10C12Q1/68
    • C12N15/1093C12Q1/6809C12Q1/6813C12Q1/6855C12Q2525/191C12Q2565/125
    • The present invention discloses a methodology which is directed to providing positive confirmation that nucleic acids, possessing putatively identified sequences predicted to generate observed GeneCalling signals, are actually present within the sample from which the signal was originally derived. The putatively identified nucleic acid fragment within the sample possesses 3'- and 5'-ends with known terminal subsequences, said method comprising: contacting said nucleic acid fragments in said sample in amplifying conditions with (i) a nucleic acid polymerase; (ii) "regular" primer oligonucleotides having sequences comprising hybridizable portions of said known terminal subsequences; and (iii) a "poisoning" oligonucleotide primer, said poisoning primer having a sequence comprising a first subsequence that is a portion of the sequence of one of said known terminal subsequences and a second subsequence that is a hybridizable portion of said putatively unidentified sequence which is adjacent to said one known terminal subsequence, wherein nucleic acids amplified with said poisoning primer are distinguishable upon detection from nucleic acids amplified with said nucleic acids amplified only with said regular primers; separating the products of the contacting step; and the detecting sequence is confirmed if the nucleic acids amplified with said poisoning primer are detected.
    • 本发明公开了一种旨在提供肯定确认的方法,核酸具有预测产生观察到的GeneCalling TM信号的推定鉴定序列实际上存在于最初得到信号的样本内。 所述样品中的推测鉴定的核酸片段具有已知末端亚序列的3'和5'末端,所述方法包括:使扩增条件下的所述样品中的所述核酸片段与(i)核酸聚合酶; (ii)具有包含所述已知末端子序列的可杂交部分的序列的“规则”引物寡核苷酸; 所述中毒引物具有包含作为所述已知末端子序列之一序列的一部分的第一子序列和作为所述假定未鉴定序列的可杂交部分的第二子序列的序列, 与所述一个已知末端子序列相邻,其中用所述中毒引物扩增的核酸在从用所述常规引物扩增的所述核酸扩增的核酸检测时是可区分的; 分离接触步骤的产物; 如果检测到用所述中毒引物扩增的核酸,则确认检测序列。