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Updated: May 12, 2025

Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection

Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection

Published on: January 13, 2014

开发一种高效的协议,从莫雷勒的鱼尾巴皮活检中提取RNA.

Asela Marisol Buenfil-Rojas^1,2, Mauricio González-Jáuregui^3,2, Mari Ochiai¹

¹Center for Marine Environmental Studies (CMES), Ehime University, 2-5 Bunkyo-cho, Matsuyama, Ehime, Japan.

|May 7, 2025

概括

我们开发了一种非侵入性RNA从鱼骨中提取RNA的方法. 该协议使得难以采样的野生物种的保护研究和转录基因分析成为可能.

关键词:

鱼类 (Crocodylians) 是一种鱼类.化组织的化组织.从莫雷勒的鱼尾巴皮切片中提取RNA 净化RNA净化RNA的过程文字转录学 (Transcriptomics) 是一个学科.

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RNA Pull-down Procedure to Identify RNA Targets of a Long Non-coding RNA

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相关实验视频

Last Updated: May 12, 2025

Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection

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Published on: January 13, 2014

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Published on: February 21, 2019

RNA Pull-down Procedure to Identify RNA Targets of a Long Non-coding RNA

RNA Pull-down Procedure to Identify RNA Targets of a Long Non-coding RNA

Published on: April 10, 2018

科学领域:

保护生物学保护生物学
分子生物学分子生物学
野生动物研究研究野生动物研究

背景情况:

从野生动物的硬组织中提取RNA具有挑战性,特别是对于需要非致命方法的保护工作.
热带地区的研究往往面临着有限的实验室基础设施,需要强大且可适应的协议.
选择了*Crocodylus moreletii*的尾状作为非侵入性RNA采样的来源.

研究的目的:

为了优化一种非侵入性RNA提取协议从*Crocodylus moreletii*的尾部.
评估RNA纯度和完整性,用于下游应用,如基因表达研究.
为其他具有硬组织的非模型物种适用的类似方法提供基础.

主要方法:

探索各种组织破坏和同质化技术.
使用RNA保存缓冲器优化RNA隔离和净化.
75毫克的样品进行超声波,然后进行3小时的蛋白酶K化.

主要成果:

获得的RNA度在18.7至154.7 ng/μL之间.
获得了令人满意的RNA纯度,其260/280比率约为2.
确认RNA完整性,RNA完整性数 (RIN) 大于5.5.
验证的RNA适用于定量PCR和下一代测序 (NGS).

结论:

优化的协议提供了一种可靠的方法,用于从鱼骨中非侵入性提取RNA.
提取的RNA适用于基因表达分析和转录学研究.
这种方法可以适用于从其他非模型野生物种的硬组织中提取RNA,从而有助于保护研究.