Invention Grant
- Patent Title: Method for amplifying a T cell receptor (TCR) cDNA
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Application No.: US15314862Application Date: 2015-05-29
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Publication No.: US10533204B2Publication Date: 2020-01-14
- Inventor: Hiroshi Hamana , Hiroyuki Kishi , Atsushi Muraguchi , Kiyomi Shitaoka
- Applicant: NATIONAL UNIVERSITY CORPORATION UNIVERSITY OF TOYAMA , SC WORLD, INC.
- Applicant Address: JP Toyama-shi JP Toyama-shi
- Assignee: National University Corporatlon University of Toyama,SC World, Inc.
- Current Assignee: National University Corporatlon University of Toyama,SC World, Inc.
- Current Assignee Address: JP Toyama-shi JP Toyama-shi
- Agency: Crowell & Moring LLP
- Priority: JP2014-113308 20140530
- International Application: PCT/JP2015/065577 WO 20150529
- International Announcement: WO2015/182749 WO 20151203
- Main IPC: C12P19/34
- IPC: C12P19/34 ; C07K14/725 ; C12Q1/6886 ; A61K48/00

Abstract:
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.
Public/Granted literature
- US20170183700A1 Method for Amplifying TCR cDNA Public/Granted day:2017-06-29
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