Dimer avoided multiplex polymerase chain reaction for amplification of multiple targets

    公开(公告)号:US20220259653A1

    公开(公告)日:2022-08-18

    申请号:US17300937

    申请日:2021-12-15

    申请人: iRepertoire, Inc.

    摘要: The present disclosure relates to methods for amplifying nucleic acids that avoid problems associated with primer-dimer formation. The present methods are referred to herein as dimer avoided multiplex polymerase chain reaction (dam-PCR). The methods disclosed herein generally comprise the steps of reverse transcribing at least one first strand of DNA, for example cDNA from an RNA sample, wherein each first strand of DNA incorporates a reverse common primer binding site; selecting each first strand of DNA; synthesizing at least one second strand of DNA from each of the at least one first strand of DNA, wherein each second strand of DNA incorporates a forward common primer binding site; selecting each second strand of cDNA; and amplifying the DNA strands using common primers. Alternatively, the method may be performed using a gDNA template. The methods described herein, due to the selection of DNA strands and removal of unused primers prior to amplification, avoid primer-dimer formation and allow for greater sensitivity and efficiency compared with conventional multiplex PCR methods.

    Immunorepertoire normality assessment method and its use

    公开(公告)号:US11047011B2

    公开(公告)日:2021-06-29

    申请号:US15278998

    申请日:2016-09-28

    申请人: iRepertoire, Inc.

    发明人: Jian Han

    IPC分类号: C12Q1/68 C12Q1/6883

    摘要: The present disclosure generally pertains to a method for comparing the immunorepertoire diversity of a patient to the shared immunorepertoire of a group of individuals to identify whether the immunorepertoire diversity of the patient is normal or abnormal relative to the shared immunorepertoire of the group of individuals.

    METHODS AND DEVICES FOR COLLECTION OF EXHALED AEROSOLS

    公开(公告)号:US20220000391A1

    公开(公告)日:2022-01-06

    申请号:US16919015

    申请日:2020-07-01

    申请人: iREPERTOIRE, INC.

    发明人: Jian Han Wenjing Pan

    IPC分类号: A61B5/097 A61B5/08 A61B10/00

    摘要: The present disclosure relates to methods and devices for the collection of exhaled aerosols. In some embodiments, the device disclosed herein comprises a cup and an attachable lid, wherein the cup has a conical base and the lid includes a pipette access port with a removable cap and a sealed buffer reservoir. In some embodiments, the method for a subject to collect exhaled aerosols disclosed herein comprises the steps of placing a collection cup against the subject's mouth, the subject coughing into the collection cup at least once, immediately closing the device by reattached a lid to the collection cup, placing the collection cup in a transport bag, and providing the transport bag to a processor. The processor can then incubate the transport bag containing the collection cup, add buffer to the collection cup to rinse and suspend any collected exhaled aerosols, and then extract the buffer containing the exhaled aerosols either for storage or for immediate amplification and sequencing for the detection of pathogens.

    SYSTEMS AND METHODS USING PIPETTES FOR PERFORMING CHEMICAL AND/OR BIOLOGICAL PROCESSES

    公开(公告)号:US20210129129A1

    公开(公告)日:2021-05-06

    申请号:US17020349

    申请日:2020-09-14

    申请人: iRepertoire, Inc.

    IPC分类号: B01L3/02

    摘要: A system for performing a chemical and/or biological process has a pipette for processing material used in the process. The pipette can have a tip that can pierce a sealing material of a chamber and then be placed in contact with the bottom surface of the chamber that is housing a material used in the process. The tip has a plurality of passageways with openings located along the outer perimeter of the tip. The placement of the openings at the outer perimeter permits the tip to be placed on the bottom surface of a chamber such that most of the material in the chamber can be extracted by the pipette. In addition, the placement of the tip on the bottom surface of the chamber with openings along the outer perimeter of the tip permits the pipette to agitate existing material in the chamber when the pipette is providing additional material into the chamber.

    DIMER AVOIDED MULTIPLEX POLYMERASE CHAIN REACTION FOR AMPLIFICATION OF MULTIPLE TARGETS

    公开(公告)号:US20200071763A1

    公开(公告)日:2020-03-05

    申请号:US16492882

    申请日:2018-03-09

    申请人: iRepertoire, Inc.

    摘要: The present disclosure relates to methods for amplifying nucleic acids that avoid problems associated with primer-dimer formation. The present methods are referred to herein as dimer avoided multiplex polymerase chain reaction (dam-PCR). The methods disclosed herein generally comprise the steps of reverse transcribing at least one first strand of DNA, for example cDNA from an RNA sample, wherein each first strand of DNA incorporates a reverse common primer binding site; selecting each first strand of DNA; synthesizing at least one second strand of DNA from each of the at least one first strand of DNA, wherein each second strand of DNA incorporates a forward common primer binding site; selecting each second strand of cDNA; and amplifying the DNA strands using common primers. Alternatively, the method may be performed using a gDNA template. The methods described herein, due to the selection of DNA strands and removal of unused primers prior to amplification, avoid primer-dimer formation and allow for greater sensitivity and efficiency compared with conventional multiplex PCR methods.