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Phenol, pH 4.3

Company: Sigma-Aldrich
Catalog#: P4682
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Total RNA Extraction from Dinoflagellate Symbiodinium Cells
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Date:
2018-06-05
[Abstract]  Dinoflagellates are unicellular algae that can have photosynthetic or nonphotosynthetic lifestyles. Dinoflagellates in the genus Symbiodinium can enter endosymbiotic associations with corals, providing the metabolic basis for the highly productive and biologically diverse coral-reef ecosystems (Hoegh-Guldberg, 1999), as well as with other cnidarians, including sea anemones and jellyfish, and non-cnidarian hosts (Trench, 1993; Lobban et al., 2002; Mordret et al., 2016).

Here, I describe a protocol for isolating total RNA from Symbiodinium cells.
[摘要]  鞭毛藻是单细胞藻类,可以有光合或非光合生活方式。 Symbiodinium属中的甲藻类可以与珊瑚进入内共生关系,为高生产力和生物多样性珊瑚礁生态系统提供代谢基础(Hoegh-Guldberg,1999),以及其他的cnidarians,包括 海葵和海蜇,以及非刺猬寄主(Trench,1993; Lobban等人,2002; Mordret等人,2016)。

在这里,我描述了从Symbiodinium细胞中分离总RNA的方案。

Polysome Analysis
Author:
Date:
2017-03-20
[Abstract]  Polysome analysis is a method to separate mRNAs from a cell into actively translating and non-translating fractions depending on their association with polysomes. By this protocol, cell lysates are fractionated by sucrose density gradient ultracentrifugation. Free mRNA fraction and various ribosomal fractions, such as 40S, 60S, monosomes and polysomes are collected by fractionation. Association of particular mRNAs with these fractions is detected by reverse transcription – PCR to investigate the translational state of the mRNA. [摘要]  多聚赖氨酸分析是一种将mRNA从细胞分离为主动翻译和非翻译部分的方法,这取决于它们与多核糖体的关联。通过该方案,通过蔗糖密度梯度超速离心分离细胞裂解物。通过分级收集游离mRNA级分和各种核糖体级分,例如40S,60S,单体和多核糖体。通过逆转录PCR检测特异性mRNA与这些级分的关联,以研究mRNA的翻译状态。

背景 细胞mRNA在任何时间点分布到主动翻译和非翻译池中,并可响应于各种刺激而在这些池之间动态地重新分布。主动翻译的mRNA具有较高数量的与它们相关的核糖体,与mRNA相关的核糖体数量是mRNA的翻译状态的量度。因此,从细胞中分离核糖体时,会在多核糖体组分中发现主要转录的mRNA,而在游离mRNA部分或与40S核糖体亚基相关的非翻译/不良翻译mRNAs。因此,多聚赖氨酸分析是根据其与多核糖体的关联将mRNA从细胞分离为主动翻译和非翻译部分的方法(Ray et al。,2009; Poria等人, >。,2016)。可以通过RT-PCR检测单个mRNA与翻译/非翻译级分的关联,而通过RNA测序或微阵列分析可以鉴定mRNA的整个翻译或非翻译池。

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