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    • 8. 发明申请
    • SCREENING FOR FUNCTIONAL ANTISENSE AGENTS
    • 筛选功能性抗菌剂
    • WO99045145A1
    • 1999-09-10
    • PCT/GB1999/000631
    • 1999-03-03
    • C12N15/09C12Q1/68
    • C12Q1/6816C12Q1/6837Y10S977/924C12Q2565/601C12Q2563/173C12Q2525/207C12Q2565/515
    • Provided is a method for identifying a functional antisense agent, which method comprises hybridising an RNA with an aligonucleotide probe and measuring in real time the kinetics of hybridisation, wherein the kinetics are measured by either hybridising in the presence of an intercalation dye and recording a change in the spectroscopic properties of the dye as hybridising proceeds, or incorporating a label in either the RNA or the probe, attaching the non-labelled RNA or non-labelled probe to a solid support, generating an evanescent wave in the proximity of the non-labelled RNA or non-labelled probe and recording the increase in a signal generated by interaction of the evanescent wave with the label, as hybridisation proceeds.
    • 提供了用于鉴定功能性反义剂的方法,该方法包括将RNA与寡核苷酸探针杂交并实时测量杂交的动力学,其中通过在插入染料存在下杂交并记录变化来测量动力学 在作为杂交进行的染料的光谱性质中,或在RNA或探针中掺入标记,将未标记的RNA或未标记的探针附着到固体支持物上, 标记的RNA或非标记探针,并记录由ev逝波与标记相互作用产生的信号的增加,因为杂交进行。
    • 10. 发明申请
    • HIGH SURFACE DENSITY COVALENT IMMOBILIZATION OF OLIGONUCLEOTIDE MONOLAYERS
    • 高表面密度共价键固定寡核苷酸单链的方法
    • WO99020640A2
    • 1999-04-29
    • PCT/CA1998/000969
    • 1998-10-16
    • G01N33/53B01J20/32B32B17/10C03C17/30C03C17/34C07C309/39C07F7/12C07H21/00C07H21/04C12M1/00C12N15/09G01N37/00
    • B82Y30/00B01J20/3242B32B17/10688B82Y5/00B82Y15/00C03C17/30C03C17/3405C07B2200/11C07H21/00Y10S977/702Y10S977/902Y10S977/92Y10S977/924Y10S977/958
    • Oligonucleotides and other biomolecules are immobilized in high density on solid substrates through covalent forces using either a permanent thioether bond, or a chemoselectively reversible disulfide bond to a surface thiol. Substrates which have hydroxyl groups on their surfaces can be first silanized with a trichlorosilane containing 2-20 carbon atoms in its hydrocarbon backbone, terminating in a protected thiol group. The oligonucleotides or other biomolecules are first connected to a tether consisting of a hydrocarbon or polyether chain of 2-20 units in length which terminates in a thiol group. This thiol may be further modified with a halobenzylic-bifunctional water soluble reagent which allows the conjugate to be immobilized onto the surface thiol group by a permanent thioether bond. Alternatively, the oligonucleotide-tether-thiol group can be converted to a pyridyldisulfide functionality which attaches to the surface thiol by a chemoselectively reversible disulfide bond. The permanently bound oligonucleotides are immobilized in high density compared to other types of thiol functionalized silane surface and to the avidin-biotin method.
    • 本发明涉及在固体基质上通过共价力使用永久硫醚键或化学选择性可转化为表面硫醇的二硫键的高密度寡核苷酸和其他固定生物分子。 羟基底物可以首先在它们的烃骨架上用含有2-20个碳原子的三氯硅烷在其表面上进行硅烷化,终止于受保护的硫醇基。 首先将寡核苷酸或其他生物分子结合到由长度在2和20个单位之间的烃或聚醚链组成的栓链上,所述单链以巯基终止。 然后可以用双官能卤代苄基水溶性试剂修饰该巯基,这使得可以通过永久硫醚键将缀合物固定在表面硫醇基上。 根据另一个变体,寡核苷酸 - 连接臂硫醇基团可以转化为吡啶基二硫化物功能,其通过化学选择性可逆的二硫键与表面硫醇结合。 与其他类型的官能化硫醇硅烷表面以及抗生物素蛋白 - 生物素方法相比,高密度永久结合的寡核苷酸被固定化。