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通过纳米链技术在高度降解的固定组织中量化mRNA:一项比较研究

Eros Azzalini¹, Barbara Di Stefano¹, Vincenzo Canzonieri^1,2

¹Department of Medical Sciences, University of Trieste, 34149 Trieste, Italy.

Methods and protocols

|May 28, 2024

概括

nCounter纳米链技术有效量化档案组织中的mRNA,在甲固定和Bouin两种情况下都胜过qPCR和ddPCR.

科学领域:

分子生物学分子生物学
翻译研究是翻译研究.
档案组织分析分析.

背景情况:

档案组织对于分子分析至关重要,但遭受生物分子降解.
固定在形式素和嵌入式抛 (FFPE) 组织是标准的,但布恩的溶液也在历史上被使用过.
研究历史处理的组织中的RNA完整性对研究至关重要.

研究的目的:

评估nCounter纳米链杂交,以量化降解的布温固定和嵌 (BFPE) 组织中的mRNA.
为了比较纳米链性能与定量PCR (qPCR) 和数字滴滴PCR (ddPCR).
评估FFPE和BFPE样本中的RNA量化可行性.

主要方法:

分析了16个嵌入的组织块 (8个FFPE,8个BFPE).
纳米链技术应用于300 ng的RNA样本.
通过使用360 ng模板的qPCR和ddPCR进行RNA逆转录和分析.

主要成果:

与qPCR和ddPCR相比,纳米链技术在FFPE和BFPE样本中检测目标mRNA方面表现优异.
该研究证实了纳米链的有效性,即使使用高度降解的RNA模板.
RNA模板降解仍然是一个限制,可能会影响基因表达分析结果.

结论:

nCounter Nanostring 是一个强大的工具,用于存档组织的mRNA量化,包括历史处理的BFPE样本.
虽然纳米链具有优势,但研究人员必须意识到RNA降解对基因表达数据的影响.
这项研究验证了纳米链用于利用各种档案组织样本进行翻译研究.

关键词:

在Bouin Bouin,我们可以看到.在FFPE的FFPE.这是一个RNARNARNARNARNA.降解RNA的降解RNA的降解 RT-ddPCRR 在线提供 RT-qPCRR 是一个人.n 计数器计数器

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