High-quality total RNA extraction from Magnolia sieboldii K. Koch seeds: a comparative evaluation
High-quality total RNA extraction from Magnolia sieboldii K. Koch seeds: a comparative evaluation
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从天女木兰种子中提取高质量总 RNA：比较评价
DOI：
10.1007/s11676-018-0615-8
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发表时间：
2019-02
影响因子：
3
通讯作者：
Lu Xiujun
中科院分区：
文献类型：
--
作者：
Wei Jun;Zhang Xiaolin;Hou Zhe;Lu Xiujun
天女木兰种子的特点是具有深度休眠。由于缺乏总 RNA（核糖核酸）提取方案，内部分子调节机制尚未得到研究。高质量 RNA 的提取很重要，并且可能成为植物分子生物学实验的限制因素。阐明萌发的分子调控机制需要足够的高质量总RNA。然而，含有大量次生代谢物的M. sieboldii 种子也含有难以分离RNA 的组织。通过对8种方法的评价和筛选并进一步优化，我们找到了两种简单、低成本的西氏花种子RNA提取方法。这两种方法不仅适用于西氏花种子RNA的提取，也适用于其他几种组织中RNA的提取。通过这些方法提取的总RNA适用于一般分子生物学实验，例如qRT-PCR（定量实时聚合酶链式反应）。该方案还满足转录组测序和小 RNA 测序的严格要求。这项研究为未来转录水平的研究提供了一种强有力的方法。
Magnolia sieboldii K. Koch seed is characterized with having deep dormancy. The inner molecular regulation mechanism has not been investigated because of the absence of a protocol for total RNA (ribonucleic acid) extraction. The extraction of high-quality RNA is important and can be a limiting factor in plant molecular biology experiments. Sufficient high-quality total RNA is required to elucidate the molecular regulation mechanism of germination. However, M. sieboldii seeds with large amounts of secondary metabolites also contain recalcitrant tissues for RNA isolation. We found two simple and low-cost RNA extraction methods for M. sieboldii seeds by evaluating and selecting eight types of methods and further optimizing these methods. The two methods were not only suitable for extracting M. sieboldii seed RNA but also applicable to RNAs from several other tissues. Total RNA extracted through these approaches was applicable for general molecular biology experiments such as qRT-PCR (quantitative real-time polymerase chain reactions). The protocols also meet the strict harsh requirements for transcriptome sequencing and small RNA sequencing. This study provides a powerful approach for future studies at the transcription level.
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