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Tween-20

吐温 20

Company: Sigma-Aldrich
Catalog#: P9416
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Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
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, isolated from fish samples was subjected to reverse transcription PCR amplification and the amplification products were mixed with specific oligonucleotide probe. A red test line was formed when ... [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 ...

RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
Author:
Date:
2020-07-20
[Abstract]  Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive method of mRNA stability measurement through quantitative reverse transcription PCR (RT-qPCR) after inhibition of RNA transcription by cordycepin in Arabidopsis seedlings. [摘要]  [摘要] 稳态mRNA的水平取决于转录和降解的速率。mRNA稳定性和/或降解的调节是可以显着影响mRNA水平及其生物学功能的关键因素。mRNA的稳定性可以在转录抑制后间接测量。该协议描述了通过拟南芥幼苗中的虫草素抑制RNA转录后,通过定量逆转录PCR(RT-qPCR)进行mRNA稳定性测定的快速灵敏方法。

[背景] mRNA稳定性的调控是基因表达调控的关键控制点。mRNA的稳定性对基因表达,分子和细胞表型,以及最终对植物发育,防御和其他生物过程都具有深远的影响。多种方法,例如RNA印迹分析,原位杂交,可用于测量转录抑制后的mRNA稳定性。在此协议中,我们描述了一种快速灵敏的方法,通过虫草素抑制转录后,通过RT-qPCR测量mRNA的稳定性。虫草素或3'-脱氧腺苷是腺苷类似物(参见参考文献2 )。可以将3'-脱氧腺苷掺入RNA,并由于3'位置不存在羟基部分而抑制转录延伸和RNA合成(参见参考文献6 )。我们已经成功地使用了这种方便而灵敏的方法来测量拟南芥幼苗中几种低丰度mRNA 的稳定性,包括初级microRNA转录本(Jia 等,2017)。在这里,我们用详细的实验程序和数据分析方法介绍该协议。

Immunolabeling of Maize Meiocytes
Author:
Date:
2020-04-05
[Abstract]  This protocol describes a quick method for immunolocalization of meiotic proteins in maize. It includes a fixation step that allows for long-term storage of material and provides good preservation of chromatin structure. [摘要]  [摘要] 该协议描述了一种在玉米中进行减数分裂蛋白免疫定位的快速方法。它包括一个固定步骤,可以长期保存材料并提供良好的染色质结构保存。

[背景] 卵母细胞是生殖细胞,在其中发生同源染色体配对,突触和重组的专门过程,然后染色体分离以产生单倍体核。阐明减数分裂蛋白的定位对于理解这些过程至关重要。尽管此协议可能有多种变体,但我们发现微波加热步骤对于使其可靠地工作于玉米中的免疫定位至关重要。

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