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Fabrication of nanopores with ultrashort single-walled carbon nanotubes inserted in a lipid bilayer.
刘蕾(多); Xie JN(谢佳妮); Wu HC(吴海臣); Liu, Lei; Xie, Jiani; Li, Ting; Wu, Hai-Chen.
2015
发表期刊Nature Protocols
卷号10期号:11页码:1670-1678
通讯作者吴海臣
文章类型Article
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.
学科领域Biochemistry & Molecular Biology
DOI10.1038/nprot.2015.112
收录类别SCI ; CA
WOS类目Biochemical Research Methods
WOS记录号WOS:000363472700002
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被引频次:10[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.ihep.ac.cn/handle/311005/228865
专题多学科研究中心
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GB/T 7714
Liu L,Xie JN,Wu HC,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 刘蕾.,谢佳妮.,吴海臣.,Liu, Lei.,Xie, Jiani.,...&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 刘蕾,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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