A method for extracting high-quality RNA from diverse plants for next-generation sequencing and gene expression analyses.
A method for extracting high-quality RNA from diverse plants for next-generation sequencing and gene expression analyses.
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DOI：
10.3732/apps.1300070
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发表时间：
2013-12
影响因子：
3.6
通讯作者：
Specht CD
中科院分区：
文献类型：
--
作者：
Yockteng R;Almeida AM;Yee S;Andre T;Hill C;Specht CD
·研究报告：为了研究植物中的基因表达，必须提取足够数量的高质量RNA用于随后的cDNA文库构建。野外采集的组织通常在数量和质量上受到限制，通常保存在RNAlater中。从不同保存的样品中获得足够的高质量的产量是比较生物学研究的必要条件。我们提出了一个协议，即使是最高质量的RNA提取植物组织。·方法和结果：来自苔藓、苏铁和被子植物花器官和叶的组织保存在RNAlater中或在−80°C下冷冻新鲜。进行提取并测量产率和纯度的质量。结论：该方案导致从代表维管和非维管植物的各种植物组织中提取高质量的RNA。RNA用于cDNA合成以产生用于下一代测序的文库，并用于使用定量PCR（qPCR）和半定量逆转录PCR（RT-PCR）的表达研究。
• Premise of the study: To study gene expression in plants, high-quality RNA must be extracted in quantities sufficient for subsequent cDNA library construction. Field-based collections are often limited in quantity and quality of tissue and are typically preserved in RNAlater. Obtaining sufficient and high-quality yield from variously preserved samples is essential to studies of comparative biology. We present a protocol for the extraction of high-quality RNA from even the most recalcitrant plant tissues. • Methods and Results: Tissues from mosses, cycads, and angiosperm floral organs and leaves were preserved in RNAlater or frozen fresh at −80°C. Extractions were performed and quality was measured for yield and purity. • Conclusions: This protocol results in the extraction of high-quality RNA from a variety of plant tissues representing vascular and nonvascular plants. RNA was used for cDNA synthesis to generate libraries for next-generation sequencing and for expression studies using quantitative PCR (qPCR) and semiquantitative reverse transcription PCR (RT-PCR).
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影响因子：
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作者：
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通讯作者：
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通讯作者：
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