ProtectRNA^TM RNase Inhibitor 500× Concentrate

ProtectRNA ^TM RNase抑制剂500×浓缩物

Company: Sigma-Aldrich

Catalog#: R7397

Analysis of Chromosome Condensation/Decondensation During Mitosis by EdU Incorporation in Nigella damascena L. Seedling Roots

Eugene V. Sheval

Published online: 2018-02-05

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Single Molecule RNA FISH in the Mammalian Oocyte

Author:

Denisa Jansova,

Date:

2015-12-05

[Abstract] RNA fluorescence in situ hybridization is a method to localize and measure gene expression in individual cell or tissue. Using multiple specific fluorescently labeled oligonucleotides greatly increases signal-to-noise ratio and thus enables detection of single RNA molecule. Around forty different DNA oligonucleotides designed to common RNA target and labeled with single fluorophore at 3´ terminus hybridizes with target RNA in fixed cells. We adapt this method to visualize target RNA in the mammalian oocyte. The ability to detect single transcript in the mammalian oocyte was challenging due to its large cell size. This method consists of four simple steps: fixation, permeabilization, hybridization and imaging. The protocol is adapted to this large nonattached cell to visualize ... [摘要] RNA荧光原位杂交是定位和测量个体细胞或组织中的基因表达的方法。使用多个特异性荧光标记的寡核苷酸大大增加信噪比，从而使得能够检测单个RNA分子。大约四十个不同的DNA寡核苷酸设计为常见的RNA靶标，并在3'端用单个荧光团标记，与固定细胞中的靶RNA杂交。我们适应这种方法可视化目标RNA在哺乳动物卵母细胞。在哺乳动物卵母细胞中检测单个转录物的能力由于其大的细胞大小而具有挑战性。该方法由四个简单的步骤组成：固定，预稳定化，杂交和成像。该方案适应这种大的非附着细胞以显现母体RNA。
各种荧光团的组合允许检测更多的RNA靶标。该方法可以与细胞器标记一起使用或用免疫荧光方案扩增。

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ProtectRNA^TM RNase Inhibitor 500× Concentrate

ProtectRNA ^TM RNase抑制剂500×浓缩物