Isolation of High Quality RNA from Phyllanthus emblica and Its Evaluation by Downstream Applications
Isolation of High Quality RNA from Phyllanthus emblica and Its Evaluation by Downstream Applications
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DOI：
10.1007/s12033-011-9492-5
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发表时间：
2012-11-01
影响因子：
2.6
通讯作者：
Singh, Kashmir
中科院分区：
文献类型：
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作者：
Kumar, Avneesh;Singh, Kashmir
下一代测序是广泛用于转录组谱分析的高通量技术。高质量RNA的分离是这种大规模转录组分析的先决条件。余甘子是一种重要的药用植物，含有丰富的维生素C、黄酮类化合物、多酚类化合物、单宁等代谢产物，具有独特的药用价值。高浓度的多糖和多酚等次生代谢产物是提取高质量RNA的障碍。RNA质量的任何妥协都会影响下游应用，并需要额外的清洁步骤，进一步减少RNA的数量。我们已经开发了一种从P. embilca中分离高质量RNA的方案。成功地评估了RNA的下游应用，如逆转录聚合酶链反应、cDNA末端的快速扩增、mRNA文库制备和使用HiSeq（（TM））2000测序技术的测序。该方案简单，可在4-5 h内完成。
Next generation sequencing is a high-throughput technique widely used for transcriptome profiling. Isolation of high quality RNA is a prerequisite for such large scale transcriptome analysis. Phyllanthus emblica is an important medicinal plant having high amount of metabolites like vitamin C, flavonoids, polyphenolic compounds, tannins, which are responsible for its wondered medicinal properties. High concentration of secondary metabolites like polysaccharides and polyphenols proved to be an obstacle in isolating RNA of good quality. Any compromise with quality of RNA affects the downstream applications and requires extra cleaning steps that further reduce RNA quantity. We have developed a protocol for isolation of high quality RNA from P. embilca. RNA was successfully assessed for downstream applications like reverse transcription polymerase chain reaction, rapid amplification of cDNA ends, mRNA library preparation, and sequencing using HiSeq((TM)) 2000 sequencing technology. The protocol is simple and can be completed in 4-5 h.