| RETRACTED: Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
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Author:
Date:
2020-08-05
[Abstract] Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health as well as animal health protection. In that aspect, the present protocol focuses on the development of functionalized gold nanoparticle-based lateral flow biosensor for fish nervous necrosis virus (Nodavirus) nucleic acids detection. Total viral RNA, ...
[摘要] [摘要] 纸纳米生物传感器已经成为分析导致各种疾病的兽医和人类病原体的一个极好的平台。尤其是侧向流分析或生物传感器是实验室设备和现场分析中灵敏、快速、可靠和准确分析的理想选择。病毒RNA检测对公共卫生和动物健康保护具有重要意义。在这一方面，本协议的重点是开发功能化金纳米粒子侧流生物传感器，用于鱼类神经坏死病毒（Nodavirus）核酸检测。从鱼类标本中分离出的总病毒RNA进行逆转录PCR扩增，扩增产物与特异性寡核苷酸探针混合。当有nodavirus产物存在时，形成红色检测线。这种方法对基础研究有很大的意义，因为它消除了耗时、繁琐的电泳程序的需要，并且可以在养鱼场对养鱼户进行调整。利用这种生物分析平台进行病害监测，无需耗费大量时间和成本，对水产养殖和环境安全有很大影响。
[背景] 关注点和/或现场生物分析一直是关注人类和动物福祉的研究工作的最终目标。基于纸基的传感平台具有功能化简单、重现性好、制造成本低等优点，是一种极具吸引力的分析平台。纸基分析设备已应用于小分子、蛋白质和各种核酸的分析（Parolo和Merkoçi，2013；Bahadir和Sezgintürk，2016；Jiang等人，2019）。侧流生物传感器（LFB）是一种带有干试剂的预制材料条带，通过流体样品激活。它们专为一次性一次性使用而设计，只要有足够的开/关信号（Posthuma ...
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| Micro-chromatin Immunoprecipation (μChIP) Protocol for Real-time PCR Analysis of a Limited Amount of Cells
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Author:
Date:
2016-06-20
[Abstract] Chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) is an important strategy to study gene regulation. When availability of cells is limited, however, it can be useful to focus on specific genes to investigate in depth the role of transcription factors or histone marks. Unfortunately, performing ChIP experiments to study transcription factors’ binding to DNA can be difficult when biological material is restricted. This protocol describes a robust method to perform μChIP for over-expressed or endogenous transcription factors using ~100,000 cells per ChIP experiment ...
[摘要] 染色质免疫沉淀后深层测序（ChIP-Seq）是研究基因调控的重要策略。 然而，当细胞的可用性有限时，可能有用的是专注于特定的基因深入研究转录因子或组蛋白标记的作用。 不幸的是，当生物材料受到限制时，进行ChIP实验以研究转录因子与DNA的结合可能是困难的。 该方案描述了使用约100,000个细胞/ChIP实验对过表达或内源性转录因子进行μChIP的稳健方法（Masserdotti等人，2015）。 我们还描述了优化步骤，我们认为这是协议工作的关键，可以用于进一步减少单元格的数量。
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| RNase H Polymerase-independent Cleavage Assay for Evaluation of RNase H Activity of Reverse Transcriptase Enzymes
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Author:
Date:
2015-08-20
[Abstract] The ribonuclease H (RNase H) polymerase-independent cleavage assay allows detection and quantification of RNase H activity of reverse transcriptase (RT) enzymes with a hybrid substrate formed by a fluorescein labeled RNA annealed with Dabcyl DNA (Figure 1). Here we describe a protocol that we have adapted for HIV-1 RT expressed from a p(His)6-tagged p66/p51 HIV-1HXB2 RT-prot plasmid and for RT of the prototype foamy virus (PFV RT).
Figure 1. Scheme of the principle of the experiment. ...
[摘要] 核糖核酸酶H（RNase H）聚合酶非依赖性切割测定允许用由与Dabcyl DNA退火的荧光素标记的RNA形成的杂交底物检测和定量逆转录酶（RT）酶的RNA酶H活性（图1）。在这里我们描述了一个协议，我们已适应的艾滋病毒1 RT表示从p（His）6标记的p66/p51艾滋病毒1 HXB2 RT-prot质粒和原型泡沫病毒（PFV RT）RT。
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