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    Substrates for Isolating, Reacting and Microscopically Analyzing Materials
    1.
    发明申请
    Substrates for Isolating, Reacting and Microscopically Analyzing Materials 审中-公开
    用于隔离，反应和显微分析材料的基板

    公开(公告)号：US20100311614A1

    公开(公告)日：2010-12-09

    申请号：US12815832

    申请日：2010-06-15

    Applicant: Jean I. Montagu ,  Roger Dowd ,  David Root

    Inventor： Jean I. Montagu ,  Roger Dowd ,  David Root

    IPC: C40B40/02 ,  B05D3/06 ,  B05D3/02 ,  B05D1/18 ,  C40B40/00 ,  C40B40/06 ,  C40B40/08 ,  C40B40/10

    CPC classification number: G01N33/544 ,  G01N33/54366

    Abstract: An immobilizing device for biological material comprises a rigid support (12) carrying a substrate layer (20, 20′) of polymer having biological immobilizing properties, e.g. for amino and nucleic acids. Substantially solid ultra-thin substrate layers (20′) having a thickness less than about 5 micron, preferably between about 0.1 and 0.5 micron, and microporous, ultra-thin substrate layers (20′) having a thickness less than about 5 micron, preferably less than 3 micron, 2 or 1 micron are shown, which may be segmented by isolating moats M. The substrate layer is on a microscope slide (302), round disc (122), bio-cassette, at the bottom of a well of a multiwell plate, and as a coating inside a tube. Fluorescence or luminescence intensity and geometric calibration spots (420) are shown. Reading is enhanced by the intensity calibration spots (420) to enable normalization of readings under uneven illumination conditions, as when reading by dark field, side illumination mode. The reference spots are shown being printed simultaneously with printing an array of biological spots or with the same equipment. Methods of forming layers of the device include controlled drawing from a bath of coating composition and drying, and spinning of C-D shaped substrates. Post-forming treatment is shown by corona treatment and radiation. Adherent metal oxides (14), silica-based materials and other materials are used to unite layers of the composite. In multiwell plates the oxide promotes joining of a bottom plate (95, 95′) and upper, well-defining structure (94) of dissimilar material. The oxides (14) also provide beneficial opacity to prevent light entering the glass support, for applying potential to the substrate, etc.

    Abstract translation: 用于生物材料的固定装置包括承载具有生物固定性质的聚合物的基底层（20,20'）的刚性支撑体（12），例如。 用于氨基和核酸。 具有厚度小于约5微米，优选约0.1至0.5微米的基本上固体的超薄基底层（20'）和厚度小于约5微米的微孔，超薄基底层（20'），优选地 示出了小于3微米，2或1微米，其可以通过分离护羊M分段。基底层在显微镜载玻片（302），圆盘（122），生物盒，在孔的底部 多孔板，以及管内的涂层。 显示荧光或发光强度和几何校准点（420）。 通过强度校准点（420）增强读数，以使得在不均匀照射条件下的读数正常化，如通过暗场，侧面照明模式读取。 参考点被显示为同时印刷生物斑点阵列或用相同的设备打印。 形成装置层的方法包括从涂料组合物的浴中进行控制拉伸和干燥，以及C-D形基材的纺丝。 后处理通过电晕处理和辐射显示。 使用粘附金属氧化物（14），二氧化硅基材料和其他材料来组合复合材料层。 在多孔板中，氧化物促进底板（95,95'）和不同材料的上部，良好限定结构（94）的接合。 氧化物（14）还提供有益的不透明性，以防止光进入玻璃支撑体，以将电位施加到基底等。

    Multi-well filter strip and composite assemblies
    4.
    发明授权
    Multi-well filter strip and composite assemblies 失效
    多孔过滤条和复合组件

    公开(公告)号：US4948564A

    公开(公告)日：1990-08-14

    申请号：US430812

    申请日：1989-11-02

    Applicant: David Root ,  George Lyman

    Inventor： David Root ,  George Lyman

    IPC: B01D29/01 ,  B01D61/18 ,  B01L3/00 ,  B01L9/06 ,  G01N21/03

    CPC classification number: B01L9/06 ,  B01D61/18 ,  B01L3/5025 ,  B01L3/50255 ,  B01L3/50855 ,  G01N2021/0357 ,  G01N2201/0446 ,  Y10S436/809 ,  Y10T436/255

    Abstract: A filter strip and composite assemblies for filtering microliter quantities of fluids. A linear array of wells having open top and bottom ends are connected by frangible webs in spaced-apart relation with discrete filter membranes closing the bottom ends of each well. Tabs are provided on the ends of the filter strip for holding the same and supporting the strip in a rectangular holder having alpha-numeric designations for identifying each well in a plurality of such filter strips contained within the holder. The filter strip may be used in a vacuum manifold for applying a pressure differential across the filter membrane and directing the filtrate into an aligned aperture of a closed bottom well of an array of wells held within the manifold. Alternatively, the filter strip may be used with a transfer plate for directing the filtrate from each well of the filter strip to an aligned well of a closed bottom array of wells. Another composite assembly includes a pressure manifold for applying an increased pressure above the membranes.

    Abstract translation: 用于过滤微量流体的过滤条和复合组件。 具有开放的顶部和底部的井的线性阵列通过易碎的腹板以间隔开的关系连接，离心过滤膜封闭每个孔的底端。 在过滤条的端部设置有用于保持其的突片，并将条带支撑在具有字母数字标号的矩形保持器中，用于识别容纳在保持器内的多个这种过滤条中的每个孔。 过滤条可以用在真空歧管中，用于在过滤膜上施加压力差，并将滤液引导到保持在歧管内的阱阵列的封闭的底部井的对准孔。 或者，过滤条可以与转移板一起使用，用于将滤液从过滤条的每个孔导向孔的封闭底部阵列的对准井。 另一复合组件包括用于在膜上施加增加压力的压力歧管。

    Method of producing short hairpin library
    6.
    发明申请
    Method of producing short hairpin library 审中-公开
    短发夹文库的生产方法

    公开(公告)号：US20070141594A1

    公开(公告)日：2007-06-21

    申请号：US11546019

    申请日：2006-10-11

    Applicant: Biao Luo ,  David Root ,  Xiaoping Yang ,  Gregory Hinkle

    Inventor： Biao Luo ,  David Root ,  Xiaoping Yang ,  Gregory Hinkle

    IPC: C40B40/08 ,  C40B50/06

    CPC classification number: C12N15/1093 ,  C12N15/111 ,  C12N2310/14 ,  C12N2310/53 ,  C12N2330/31

    Abstract: Described herein is a method of cloning synthetic oligos (including in situ synthesized oligos) into an (one or more) expression vector for library (e.g., shRNA library) production. The oligos are synthesized with one portion of the first stem of the hairpin, followed by a first loop sequence, the complete second stem, a second loop sequence, and finished with the remaining portion of the first stem of the hairpin. The two portions of the first stem anneal to the second stem, juxtaposing the 5′ end close to the 3′ end of the oligo. The methods described herein selected for hairpins with perfectly base-paired stems. After annealing, a ligase is added to the annealed oligos and the base-paired hairpins are preferentially annealed, and ligated, creating closed circular oligos. The now circularized hairpins served as templates for rolling circle amplification using a polymerase with high processivity. One or more primers complementary to the two strands of the amplified double stranded circular hairpins initiate the rolling circle amplification in the presence of a polymerase. Using primers (e.g., a sense and antisense primer), the rolling circle amplification yields double stranded hairpin sequences. These can be digested (e.g., using restriction enzymes) to produce a double-stranded hairpin fragment encoding a single hairpin. The fragment can be cloned into an appropriately digested vector for a variety of uses including expression.

    Abstract translation: 本文描述了将合成寡核苷酸（包括原位合成的寡核苷酸）克隆到（一个或多个）用于文库（例如shRNA文库）生产的表达载体中的方法。 寡核苷酸与发夹的第一茎的一部分合成，随后是第一环序列，完整的第二茎，第二环序列，并用发夹的第一茎的剩余部分完成。 第一茎的两个部分与第二茎退火，将5'末端与寡核苷酸的3'末端并列。 本文所述的方法选择用于具有完全碱基配对茎的发夹。 退火后，将连接酶加入到退火的寡核苷酸中，并且将碱基配对的发夹优先退火并连接，产生闭合的环状寡核苷酸。 现在的圆形发夹作为使用高活性的聚合酶进行滚环扩增的模板。 与扩增的双链环状发夹的两条链互补的一个或多个引物在聚合酶存在下引发滚环扩增。 使用引物（例如有义和反义引物），滚环扩增产生双链发夹序列。 这些可以被消化（例如，使用限制酶）以产生编码单个发夹的双链发夹片段。 可以将片段克隆到适当消化的载体中，用于多种用途，包括表达。

    Culture device having a detachable cell or tissue growth surface
    9.
    发明授权
    Culture device having a detachable cell or tissue growth surface 失效
    具有可分离细胞或组织生长表面的培养装置

    公开(公告)号：US5139951A

    公开(公告)日：1992-08-18

    申请号：US595305

    申请日：1990-10-10

    Applicant: David Butz ,  George Lyman ,  David Root ,  Gregory Mathus

    Inventor： David Butz ,  George Lyman ,  David Root ,  Gregory Mathus

    IPC: C12M3/06

    CPC classification number: C12M25/02 ,  C12M23/00 ,  C12M23/12

    Abstract: A tissue or cell growth device is described for placement within a well of a cluster dish. The device has a cell or tissue retention element detachably attached to a hanger. The hanger preferably has openings allowing access to the well without removal of the member and is capable of positioning the cell or tissue retention element a preselected distance relative to the hanger. Also described are various embodiments of the device having differently shaped hangers and the combination of the device and a cluster dish.

    Abstract translation: 描述了组织或细胞生长装置用于放置在簇盘的孔内。 该装置具有可拆卸地附接到衣架的细胞或组织保持元件。 衣架优选地具有允许进入井的开口，而不移除构件，并且能够将电池或组织保持元件相对于衣架定位成预选的距离。 还描述了具有不同形状的悬挂器以及装置和簇盘的组合的装置的各种实施例。

    Apparatus for growing tissue cultures in vitro
    10.
    发明授权
    Apparatus for growing tissue cultures in vitro 失效
    用于体外培养组织培养物的装置

    公开(公告)号：US5026649A

    公开(公告)日：1991-06-25

    申请号：US841562

    申请日：1986-03-20

    Applicant: George Lyman ,  Gregory Mathus ,  David Root

    Inventor： George Lyman ,  Gregory Mathus ,  David Root

    IPC: C12M1/20 ,  C12M1/22 ,  C12M3/00 ,  C12M3/06

    CPC classification number: C12M29/04 ,  C12M25/04

    Abstract: Apparatus for growing tissue cultures in vitro, which permits a concentration gradient of nutrients to develop through a permeable membrane to which a sample of tissue is attached. The permeable membrane is attached to the bottom end of a tubular support that in turn hangs by a flange connected to its upper end on the top of a well containing the nutrients. Typically, the well is part of a tissue culture cluster dish. The flange of the support positions the support and membrane centrally in the well so as to avoid capillary action in the space between the well and support. The configuration of the support and its cooperation with the lid of the cluster dish also prevents the support and membrane from floating in the nutrient solution in the well. Openings in the support provide access for a pipette to add and withdraw fluid from the space between the well and membrane support and from the space below the membrane.

    Abstract translation: 用于在体外生长组织培养物的装置，其允许营养物的浓度梯度通过附着有组织样品的渗透膜发育。 可渗透膜附接到管状支撑件的底端，管状支撑件的底端又通过连接到容纳营养物质的孔的顶部上方的凸缘而悬挂。 通常，孔是组织培养簇盘的一部分。 支撑件的凸缘将支撑件和膜中心定位在井中，以避免在井和支撑件之间的空间中的毛细作用。 支架的配置及其与簇盘盖的配合也可防止支撑和膜浮在井中的营养液中。 支架中的开口提供移液管的通道，以从孔和膜支撑之间的空间以及膜下方的空间添加和抽出流体。

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