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    • 8. 发明授权
    • Devices and methods for the synthesis of nucleic acids
    • 用于合成核酸的装置和方法
    • US07691316B2
    • 2010-04-06
    • US10776694
    • 2004-02-12
    • Nam NgoLaurent JaquinodHong Wang
    • Nam NgoLaurent JaquinodHong Wang
    • B29C39/36B29C67/04
    • B01L3/50255B01L2200/12B01L2300/0829
    • Cylindrical devices (frits) are prepared by embedding aminoalkyl- or mercaptoalkyl-modified Controlled Pore Glass (CPG) in high-density polyethylene. Methods and devices pertaining to their use in the synthesis of nucleic acids are described. A reusable synthesis column or a reusable 96-chamber synthesis plate have been designed to hold one to 96 of the said frits that are inserted reproducibly into the synthesis chambers with a frit insertor. A short gas surpressure is required to drive entry of chemical reagents into the said frit. Reagents are retained into the frit until a second, longer surpressure is applied to drain the said reagents.
    • 通过将氨基烷基或巯基烷基改性的控制孔玻璃(CPG)嵌入高密度聚乙烯中来制备圆柱形装置(玻璃料)。 描述了它们在核酸合成中的用途的方法和装置。 已经设计了可重复使用的合成柱或可重复使用的96室合成板,以容纳一至96个所述玻璃料,其被再次插入具有玻璃料插入物的合成室中。 需要短时间的气体压力来驱动化学试剂进入所述玻璃料。 将试剂保留在玻璃料中,直到施加第二次较长的耐压以排出所述试剂。
    • 10. 发明申请
    • Multi layer chromatography of nucleic acids
    • 核酸多层色谱法
    • US20080033158A1
    • 2008-02-07
    • US11888490
    • 2007-07-31
    • Nam NgoLaurent Jaquinod
    • Nam NgoLaurent Jaquinod
    • C07H21/04
    • C12N15/101
    • Herein, we introduce three multimodal SPE methods using two to (n) purification columns to separate full length 5′-DMT-on oligonucleotides with size ranging from 40 to 180-mers from short length 5′-DMT-on oligonucleotides. Two of the said methods require using some columns sequentially with the collection and reprocessing of an intermediate fraction and are used for oligonucleotides with length ranging from 70 to 180-mers. A third method is carried out with columns stacked and used in series and is best used to purify oligonucleotides with length ranging from 40 to 80-mers. Preferentially, a series of stacked columns contains from top to bottom hydrophobic porous sorbents with increasing pore sizes. Short length DMT-on oligonucleotides arise from depurination or branching during phosphoramidite based synthesis. Reversed phase partitioning and binding of short length DMT-on oligonucleotides take place simultaneously with the size exclusion of the full length DMT-on oligonucleotides. In the presence of a high ionic strength buffer, the short length DMT-on oligonucleotides bind to the top stacked columns while the less hydrophobic contaminant or DMT-off failures do not bind and/or are being washed off. In a stacked configuration, the full length DMT-on oligonucleotides are retained by the bottom column while in a sequential configuration, full length DMT-on oligonucleotides are collected and reprocessed. After detritylation of the full length oligonucleotides from the bottom column or last column in a sequence, full length nucleic acids are eluted with purity typically ranging from 90 to 95% for oligonucleotides about 80-mers in size and purity around 80 to 90% for oligonucleotides about 150-mers in size. This invention yields purified long oligonucleotides at a fraction of traditional purification costs which could spur their wider use in biological applications.
    • 在这里,我们引入三种多模态SPE方法,使用两个到(n)纯化柱来分离大小范围在从40-180毫米的短长度5'-DMT-寡核苷酸的全长5'-DMT-寡核苷酸。 所述方法中的两个需要依次使用一些列进行中间馏分的收集和重新处理,并且用于长度范围为70-180的寡核苷酸。 第三种方法用柱堆叠并串联使用,最适用于纯化长度范围为40至80毫升的寡核苷酸。 优选地,一系列堆叠的柱含有从上到下的具有增加的孔径的疏水性多孔吸附剂。 短的DMT-寡核苷酸产生于基于亚磷酰胺的合成期间的去除或分支。 反转相分配和短长度DMT-寡核苷酸的结合与全长DMT-寡核苷酸的大小排除同时进行。 在存在高离子强度缓冲液的情况下,短长度DMT-on寡核苷酸结合顶部堆叠柱,而疏水性较小的污染物或DMT脱离失败不结合和/或被洗掉。 在堆叠配置中,全长DMT-on寡核苷酸由底部柱保留,而在顺序配置中,全长DMT-寡核苷酸被收集并再处理。 在序列中的底部柱或最后一列的全长寡核苷酸脱结合后,对于寡核苷酸,对于寡核苷酸,对于寡核苷酸,大约80个多聚体的寡核苷酸的纯度通常为90-95%,对于寡核苷酸,纯度约为80-90%,全长核酸 大约150米。 本发明以传统净化成本的一小部分产生纯化的长寡核苷酸,这可以促进其在生物应用中的更广泛的应用。