利索-全球专利检索

  1. 登录
  2. 注册

搜索结果

2 条记录 (耗时 0.015 秒)

    TCR cDNA AMPLIFICATION METHOD
    2.
    发明公开
    TCR cDNA AMPLIFICATION METHOD 审中-公开
    TCR cDNA扩增方法

    公开(公告)号：EP3165605A1

    公开(公告)日：2017-05-10

    申请号：EP15798675.3

    申请日：2015-05-29

    申请人： National University Corporation University Of Toyama ,  SC World Inc.

    发明人： HAMANA Hiroshi ,  KISHI Hiroyuki ,  MURAGUCHI Atsushi ,  SHITAOKA Kiyomi

    IPC分类号： C12N15/09 ,  C12Q1/68

    摘要： As a method for highly efficiently amplifying a TCR cDNA in a short period of time, there is provided a method for amplifying a T cell receptor (TCR) cDNA, which comprises the following step (1) and step (2):
    (1) the step of performing PCR by using at least one kind of the L primer mentioned below, the C primer 1 or UTR primer 1 mentioned below, and cDNA obtained from a single cell as the template to obtain an amplification product 1;
    - an L primer of 30- to 60-nucleotide length comprising an adapter part of 15- to 25-nucleotide length, and a leader region-annealing part of 15- to 25-nucleotide length, which is ligated downstream from the adapter part, and can anneal to a part of a leader region containing a translation initiation codon, or an upstream part thereof,
    - a C primer 1 of 15- to 25-nucleotide length, which can anneal to a part of a constant region, or a UTR primer 1 of 15- to 25-nucleotide length, which can anneal to a part of a 3' untranslated region;

    (2) the step of performing PCR by using the adaptor primer mentioned below, the C primer 2 or UTR primer 2 mentioned below, and the amplification product 1 as the template to obtain an amplification product 2;
    - an adapter primer of 15- to 25-nucleotide length, which can anneal to the adapter part of the amplification product 1,
    - a C primer 2 of 15- to 25-nucleotide length, which can anneal to a part of the constant region existing upstream from the region to which the C primer 1 anneals, or a UTR primer 2 of 15- to 25-nucleotide length, which can anneal to a part of the 3' untranslated region existing upstream from the region to which the UTR primer 1 anneals.

    摘要翻译： 作为在短时间内高效扩增TCR cDNA的方法，提供了一种扩增T细胞受体（TCR）cDNA的方法，其包括以下步骤（1）和步骤（2）：（1） 通过使用至少一种下述L引物，下述C引物1或UTR引物1和从单细胞获得的cDNA作为模板进行PCR以获得扩增产物1的步骤; - 包含15至25个核苷酸长度的衔接头部分和15至25个核苷酸长度的前导区退火部分的30至60个核苷酸长度的L引物，其连接于衔接头部分的下游， 并且可以与含有翻译起始密码子的前导区的一部分或其上游部分退火， - 可以退火至恒定区的一部分的15至25个核苷酸长度的C引物1，或UTR 15至25个核苷酸长度的引物1，其可以与3'非翻译区的一部分退火; （2）使用后述的接合引物，后述的C引物2或UTR引物2，扩增产物1作为模板进行PCR，得到扩增产物2的步骤; - 可以与扩增产物1的接头部分退火的15至25个核苷酸长度的接头引物， - 15至25个核苷酸长度的C引物2，其可以退火至恒定区的一部分 存在于C引物1退火的区域的上游或15-25个核苷酸长度的UTR引物2，所述UTR引物2可退火至存在于UTR引物1所在的区域上游的3'非翻译区的一部分 退火。