PLASMID STANDARD FOR USE IN QUANTITATIVE ASSAYS USING FLUORESCENT QUANTITATIVE PCR
Номер патента: US20130217115A1. Автор: Chen Zhao, LI JUN, MO Minli, Xu Junpu. Владелец: . Дата публикации: 22-08-2013.Disclosed is a plasmid standard for use in fluorescent quantitative PCR assays. More specifically, the present invention provides a plasmid standard as well as amplification primers and detection probes thereof for use in the detection of gene mutation and expression amount. 1. A plasmid standard for detection by fluorescent quantitative PCR , characterized in that the vector for said plasmid is pMD18-T , and the insert for said plasmid is any one of following (1)-(4) sequence to be detected , which is for detecting the mutation of corresponding gene ,(1) EGFR Exon 18, 19, 21;(2) KRAS Codon 12, 13;(3) BCRP position 482 amino acid; or(4) BRAF position 600 amino acid.2. A plasmid standard according to claim 1 , characterized in that the insert for said plasmid is:(1) SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216;(2) SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221, SEQ ID NO: 222;(3) SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225; or(4) SEQ ID NO: 226, SEQ ID NO: 227.3. A plasmid standard according to claim 1 , characterized in that the primer for preparing and detecting said plasmid is:(1) SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23;(2) SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38;(3) SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49; or(4) SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56. ...