公开(公告)号：WO2012139125A2

公开(公告)日：2012-10-11

申请号：PCT/US2012/032786

申请日：2012-04-09

Applicant: LIFE TECHNOLOGIES CORPORATION ,  SCHULTZ, Jonathan ,  NOBILE, John ,  REED, Brian ,  THWAR, Prasanna ,  ROSWECH, Todd

Inventor： SCHULTZ, Jonathan ,  NOBILE, John ,  REED, Brian ,  THWAR, Prasanna ,  ROSWECH, Todd

IPC: B01F3/08 ,  B01F5/04 ,  B01F13/00 ,  C12Q1/68 ,  B04B5/04 ,  B04B7/02 ,  B01L3/00 ,  B01L7/00

CPC classification number: B01F5/0602 ,  B01F3/0807 ,  B01F5/0485 ,  B01F13/0059 ,  B01F2215/0037 ,  B01L3/502784 ,  B04B5/0414 ,  B04B7/02

Abstract: An automated on-touch template bead preparation system is provided and includes a membrane-based emulsion generation subsystems, an emulsion PCR (ePCR) thermocycling plate and subsystem, and a continuous centrifugation emulsion breaking and templated bead collection subsystem. The emulsion generation subsystem provides uniformity in the preparation of an inverse emulsion and may be used to create large or small volume inverse emulsions rapidly and reproducibly. An emulsion-generating device is provided that can supply a continuous stream of an inverse emulsion to a thermocycling subsystem, in automated fashion. The ePCR subsystem can continuously thermocycle an inverse emulsion passed therethrough and includes static temperature zones and a consumable thermocycling plate. The continuous centrifugation subsystem can continuously break a thermally cycled inverse emulsion and collect template beads formed in the aqueous microreactor droplets of the inverse emulsion.

Abstract translation: 提供了一种自动化的在线模板珠粒制备系统，包括膜基乳液生成子系统，乳液PCR（ePCR）热循环板和子系统，以及连续离心乳液破乳和模板珠收集子系统。 乳液生成子系统在制备反相乳液时提供均匀性，并可用于快速且可再现地产生大体积或小体积的反相乳液。 提供一种乳液生成装置，其能够以自动方式向逆循环子系统提供连续的反相乳液流。 ePCR子系统可以连续热循环通过的反相乳液，并包括静态温度区和可消耗的热循环板。 连续离心子系统可以连续地破坏热循环的反相乳液并收集形成在反相乳液的微量微反应器液滴中的模板珠粒。

公开(公告)号：WO2018071522A1

公开(公告)日：2018-04-19

申请号：PCT/US2017/056113

申请日：2017-10-11

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： LIGHT, David ,  CHEN, Shann-Ching ,  DHINGRA, Dalia ,  TOM, Warren ,  ROSENBAUM, Abraham ,  SEEGER, Collyn ,  THWAR, Prasanna

IPC: C12Q1/68 ,  C12Q1/6853 ,  C12Q1/6865

CPC classification number: C12Q1/6846 ,  C12Q1/6853 ,  C12Q1/6865 ,  C12Q2525/149 ,  C12Q2563/149 ,  C12Q2565/537 ,  C12Q2525/191 ,  C12Q2525/161

Abstract: In some embodiments, the disclosure relates generally to methods, and related compositions, systems, kits and apparatuses, for conducting a stream-lined nucleic acid library preparation workflow that generally includes binding a polynucleotide (from an initial nucleic acid sample) directly to capture primers that are immobilized on a support (e.g., beads or flowcell), and amplifying the bound polynucleotide to produce a support that carries a substantially monoclonal population of the polynucleotide (or a portion thereof). The support that carries the substantially monoclonal population of the polynucleotide can be generated, using the present teachings, without performing a separate adaptor-joining step and is ready for sequencing.

Abstract translation: 在一些实施方案中，本公开内容一般涉及用于进行流线型核酸文库制备工作流程的方法和相关组合物，系统，试剂盒和设备，其通常包括结合多核苷酸（来自初始 核酸样品）直接捕获固定在载体（例如珠或流动池）上的引物，并扩增结合的多核苷酸以产生携带基本上单克隆群体的多核苷酸（或其一部分）的载体。 使用本发明的教导，可以产生携带基本上单克隆的多核苷酸群体的支持物，而无需执行单独的衔接子连接步骤并准备好测序。