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    • 9. 发明公开
    • GENE-SPECIFIC NON-BIASED AMPLIFICATION METHOD
    • 基因特异性非偏置扩增方法
    • EP3263716A1
    • 2018-01-03
    • EP16755460.9
    • 2016-02-23
    • Tohoku University
    • OGASAWARA Koetsu
    • C12Q1/68C12N15/09
    • The present invention is intended to provide a method of amplifying a target gene without bias and an adapter DNA use therefor. The adapter DNA of the present invention is double-stranded adapter DNA, which is used for unbiased gene amplification and has the following features: (a) the double-stranded adapter DNA has a sense strand and an antisense strand that are annealed with each other, the base length of the sense strand being equal to or longer than the base length of the antisense strand; (b) the base length of the sense strand is 15 to 40 bp; (c) the antisense strand includes a plurality of uracil bases, each uracil is removed by treating an adapter portion with uracil DNA glycosylase (UNG), and then, heat treatment is performed to degrade the antisense strand; (d) at least one end of the adapter DNA is in the form of a blunt end; (e) the other end of the adapter DNA binds to a target gene to be amplified; and (f) a part or all of the sense strand corresponds to a forward primer sequence used for gene amplification.
    • 本发明旨在提供一种无偏倚地扩增靶基因的方法以及为此使用的适配器DNA。 本发明的接头DNA是用于无偏基因扩增的双链接头DNA,具有以下特征:(a)双链接头DNA具有彼此退火的有义链和反义链 ,有义链的碱基长度等于或长于反义链的碱基长度; (b)有义链的碱基长度为15至40bp; (c)反义链包括多个尿嘧啶碱基,通过用尿嘧啶DNA糖基化酶(UNG)处理衔接子部分除去每个尿嘧啶,然后进行热处理以降解反义链; (d)衔接头DNA的至少一个末端为钝端形式; (e)衔接头DNA的另一端结合待扩增的靶基因; 和(f)有义链的一部分或全部对应于用于基因扩增的正向引物序列。