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	Fabrication of nanopores with ultrashort single-walled carbon nanotubes inserted in a lipid bilayer
	Liu, Lei; Xie, Jiani; Li, Ting; Wu, Hai-Chen
刊名	Nature protocols
	2015-11-01
卷号	10期号:11页码:1670-1678
ISSN号	1754-2189
DOI	10.1038/nprot.2015.112
通讯作者	Wu, hai-chen(haichenwu@ihep.ac.cn)
英文摘要	We describe a protocol for the insertion of ultrashort single-walled carbon nanotubes (swcntcntcnts) to form nanopores in a montal-mueller lipid bilayer. the swcntcntcnts are designed to bind to a specific analyte of interest; binding will result in the reduction of current in single-channel recording experiments. the first stage of the proceproceproceproceprocedureureure is to cut and separate the swcntcntcnts. we cut long, purified swcntcntcnts with sonication in concentrated sulfuric acid/nitric acid (3/1). isolation of ultrashort swcntcntcnts is carried out by size-exclusion hplcplcplc separation. the second stage is to insert these short swcntcntcnts into the lipid bilayer. this step requires a microinjection probe made from a glass capillary. the setup for protein nanopore research can be adopted for the single-channel recording experiments without any special treatment. the obtained current traces are of very high quality, showing stable baselines and little background noise. example procedures are shown for investigating ion transport and dnana translocation through these swcntcntcnt nanopores. this nanopore has potential applications in molecular sensing, nanopore dnana sequencing and early disease diagnosis. for example, we have selectively detected modified 5-hydroxymethylcytosine in single-stranded dnana (ssdnana), which may have implications in screening specific genomic dnana sequences. the protocol takes similar to 15 d, including swcntcntcnt purification, cutting and separation, as well as the formation of swcntcntcnt nanopores for dnana analyses.
WOS关键词	SOLID-STATE NANOPORES ; ALPHA-HEMOLYSIN NANOPORE ; ION-CHANNEL ; PROTEIN NANOPORES ; PEPTIDE NANOTUBES ; STRANDED-DNA ; MEMBRANE ; TRANSLOCATION ; TRANSPORT ; PORE
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemical Research Methods
语种	英语
出版者	NATURE PUBLISHING GROUP
WOS记录号	WOS:000363472700002
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2176489
专题	高能物理研究所
通讯作者	Wu, Hai-Chen
作者单位	Chinese Acad Sci, Inst High Energy Phys, Key Lab Biomed Effects Nanomat & Nanosafety, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Liu, Lei,Xie, Jiani,Li, Ting,et al. Fabrication of nanopores with ultrashort single-walled carbon nanotubes inserted in a lipid bilayer[J]. Nature protocols,2015,10(11):1670-1678.
APA	Liu, Lei,Xie, Jiani,Li, Ting,&Wu, Hai-Chen.(2015).Fabrication of nanopores with ultrashort single-walled carbon nanotubes inserted in a lipid bilayer.Nature protocols,10(11),1670-1678.
MLA	Liu, Lei,et al."Fabrication of nanopores with ultrashort single-walled carbon nanotubes inserted in a lipid bilayer".Nature protocols 10.11(2015):1670-1678.

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