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相关概念视频

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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DNA Microarrays02:34

DNA Microarrays

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

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基于纳米探针的光学编码多重复合策略用于MicroRNA分析

Sungi Kim1, Jeong-Eun Park1, Woosung Hwang1

  • 1Department of Chemistry, Seoul National University , Seoul 08826, South Korea.

Journal of the American Chemical Society
|February 10, 2017
PubMed
概括

这项研究引入了一种新的多重分子检测策略,使用光动力学编码的纳米探针进行同时定量微RNA分析. 该测试可快速和精确地检测多个microRNA目标,有助于癌症诊断.

科学领域:

  • 生物技术
  • 纳米技术
  • 分子生物学

背景情况:

  • 对相互作用分子的多重分析对于理解生物系统至关重要,特别是在细胞生物学,生物传感和癌症诊断中进行微RNA (miRNA) 分析.
  • 准确的癌症诊断需要同时检测多个miRNA序列.

研究的目的:

  • 开发一个多重分子检测策略,以可靠和定量地对多个microRNA目标进行分析.
  • 展示开发的试验在癌症诊断中的潜在应用能力.

主要方法:

  • 使用光动力学 (OK) 编码的纳米探针 (NP) 具有明确的光学信号 (红色,绿色,蓝色),与支持的脂质双层 (SLB) 连接.
  • 使用暗场显微镜 (DFM) 进行现场单颗粒监测和对组合NP组件进行规范化的RGB分析.
  • 开发了OK-nanoprobe-lipid双层 (OK-NLB) 试验,用于区分和量化多个miRNA目标.

主要成果:

  • 在单个样本中,OK-NLB测定成功区分和量化了9个不同的miRNA目标.
  • 在1小时内实现多个miRNA目标的同时检测,具有高量化和特异性.
  • 使用单基不匹配实验和HeLa细胞提取的总RNA验证了试验结果,结果与定量逆转录聚合酶链反应 (qRT-PCR) 相似.

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结论:

  • 基于OK-NP的测定为高灵敏度和特异性的多重分子检测提供了强大的工具.
  • 这种测试对推进基于miRNA的诊断具有重大潜力,特别是在早期癌症检测方面.