Improvement of multiplex semi-nested PCR system for screening of rare mutations by high-throughput sequencing

doi:10.2144/btn-2019-0001

CORC > 植物研究所 > 中国科学院植物研究所 > 中科院植物分子生理学重点实验室

	Improvement of multiplex semi-nested PCR system for screening of rare mutations by high-throughput sequencing
	Zhang, Yingchun 2; Chi, Xu 4; Feng, Laibao 3; Wu, Xingwen 2; Qi, Xiaoquan 2
刊名	BIOTECHNIQUES
	2019
卷号	67 期号:6 页码:294-298
关键词	dilution improvement multiplexed target enrichment mutants next generation sequencing
ISSN号	0736-6205
DOI	10.2144/btn-2019-0001
文献子类	Article
英文摘要	The CRISPR/Cas9 system is an efficient gene-editing method, but it is difficult to obtain mutants for some specific species and special genome structures. A previously reported multiplexed, semi-nested PCR target-enrichment approach, which does not rely on transgenic technology, has been shown to be an effective and affordable strategy for the discovery of rare mutations in a large sodium azide-induced rice population. However, this strategy has the potential for further optimization. Here, we describe an improved multiplex semi-nested PCR target-enrichment strategy with simplified processing procedures, reduced false-positive rates and increased mutation detection frequency (1 mutation/73 Kb).
学科主题	Biochemical Research Methods ; Biochemistry & Molecular Biology
电子版国际标准刊号	1940-9818
出版地	LONDON
WOS关键词	REVERSE ; RNA ; ENDONUCLEASES ; ARABIDOPSIS ; DISCOVERY ; WHEAT
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	FUTURE SCI LTD
WOS记录号	WOS:000507374000008
资助机构	National Transgenic Megaproject of China [2016ZX08009001-006]
内容类型	期刊论文
源URL	[http://ir.ibcas.ac.cn/handle/2S10CLM1/19565]
专题	中科院植物分子生理学重点实验室
作者单位	1.Chinese Acad Sci, Beijing Inst Genom, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China 3.Chinese Acad Sci, Innovat Acad Seed Design, Beijing 100101, Peoples R China 4.Univ Chinese Acad Sci, 19 A Yuquan Rd, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Yingchun,Chi, Xu,Feng, Laibao,et al. Improvement of multiplex semi-nested PCR system for screening of rare mutations by high-throughput sequencing[J]. BIOTECHNIQUES,2019,67(6):294-298.
APA	Zhang, Yingchun,Chi, Xu,Feng, Laibao,Wu, Xingwen,&Qi, Xiaoquan.(2019).Improvement of multiplex semi-nested PCR system for screening of rare mutations by high-throughput sequencing.BIOTECHNIQUES,67(6),294-298.
MLA	Zhang, Yingchun,et al."Improvement of multiplex semi-nested PCR system for screening of rare mutations by high-throughput sequencing".BIOTECHNIQUES 67.6(2019):294-298.