条形码
核酸
寡核苷酸
分析物
核酸检测
检出限
核酸法
DNA
分子生物学
化学
纳米技术
生物
色谱法
组合化学
生物化学
材料科学
计算机科学
操作系统
作者
Haley D. Hill,Chad A. Mirkin
出处
期刊:Nature Protocols
[Springer Nature]
日期:2006-06-01
卷期号:1 (1): 324-336
被引量:533
标识
DOI:10.1038/nprot.2006.51
摘要
The recently developed bio-barcode assay for the detection of nucleic acid and protein targets without PCR has been shown to be extraordinarily sensitive, showing high sensitivity for both nucleic acid and protein targets. Two types of particles are used in the assay: (i) a magnetic microparticle with recognition elements for the target of interest; and (ii) a gold nanoparticle (Au-NP) with a second recognition agent (which can form a sandwich around the target in conjunction with the magnetic particle) and hundreds of thiolated single-strand oligonucleotide barcodes. After reaction with the analyte, a magnetic field is used to localize and collect the sandwich structures, and a DTT solution at elevated temperature is used to release the barcode strands. The barcode strands can be identified on a microarray via scanometric detection or in situ if the barcodes carry with them a detectable marker. The recent modification to the original bio-barcode assay method, utilizing DTT, has streamlined and simplified probe preparation and greatly enhanced the quantitative capabilities of the assay. Here we report the detailed methods for performing the ligand exchange bio-barcode assay for both nucleic acid and protein detection. In total, reagent synthesis, probe preparation and detection require 4 d.
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