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Agilent 2100 Bioanalyzer System

Agilent 2100生物分析仪系统

Company: Agilent Technologies
Catalog#: Agilent 2100
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RNA Purification from the Unicellular Green Alga, Chromochloris zofingiensis
Author:
Date:
2018-04-05
[Abstract]  Chromochloris zofingiensis is a unicellular green alga that is an emerging model species for studies in fields such as biofuel production, ketocarotenoid biosynthesis and metabolism. The recent availability of a high-quality genome assembly facilitates systems-level analysis, such as RNA-Seq. However, cells of this alga have a tough cell wall, which is a challenge for RNA purification. This protocol was designed specifically to breach the cell wall and isolate high-quality RNA suitable for RNA-Seq studies. [摘要]  Chromochloris zofingiensis 是一种单细胞绿藻,是生物燃料生产,酮类胡萝卜素生物合成和代谢等领域研究的新兴模式物种。 最近获得的高质量基因组组装便于系统级分析,如RNA-Seq。 然而,这种藻类的细胞具有坚韧的细胞壁,这对于RNA纯化来说是一个挑战。 该方案专门设计用于破坏细胞壁并分离适合RNA-Seq研究的高质量RNA。

【背景】Chromochloris zofingiensis 是一种来自叶绿体谱系的小型单细胞绿藻(Dönz,1934)。此前,该物种在文献中已被描述为属于Muriella ,小球藻和 Mychonastes (Fučíková和Lewis,2012)。 C中有强烈的经济利益。因为它能够生产大量用于生物燃料的脂质,并且显示有望成为具有商业价值的营养制品虾青素的来源(Breuer等人,2012; Mulders等人。,2014; Liu et。,2016)。最近,发布了高质量的染色体水平的基因组组装和附带的注释,这有助于系统水平分析,如RNA-Seq(Roth等人,2017)。以下方案被设计为从液体培养物中产生高度纯化的总RNA,包括miRNA。 zofingiensis 适合RNA-Seq。 ℃。 zofingiensis 细胞受坚固细胞壁的保护,该细胞壁被设计为穿透。该方案的起始材料可以高达2.5×10 ...

In situ Hybridization (ISH) in Preparasitic and Parasitic Stages of the Plant-parasitic Nematode Meloidogyne spp.
Author:
Date:
2018-03-20
[Abstract]  The spatio-temporal expression pattern of a gene provides important indications to better understand its biological function. In situ hybridization (ISH) uses a labeled complementary single-stranded RNA or DNA probe to localize gene transcripts in a whole organism, a whole organ or a section of tissue. We adapted the ISH technique to the plant parasite Meloidogyne spp. (root-knot nematode) to visualize RNAs both in free-living preparasitic juveniles and in parasitic stages settled in the plant tissues. We describe each step of the probe synthesis, digoxigenin (DIG) labeling, nematode extraction from plant tissue, and ISH procedure. [摘要]  基因的时空表达模式为更好地理解其生物学功能提供了重要的指示。 原位杂交(ISH)使用标记的互补单链RNA或DNA探针来定位整个生物体,整个器官或一部分组织中的基因转录物。 我们将ISH技术应用于植物寄生虫

【背景】到目前为止,植物寄生性线虫的稳定转化尚未成功。 ISH能够在整个装载的Meloidogyne spp中分析体内时空基因表达。线虫。这些根结线虫在土壤中以微小蚓状幼虫(J2)形式孵化并感染宿主植物根部。 J2s穿透根部并迁移到根部维管柱状细胞。幼虫定居在根部,发育成J3和J4寄生幼鱼,诱导分化专化饲养细胞。线虫最终发育成梨形雌性,将在根表面释放数百个卵。在这里,我们报告了一个详细的协议来检测准备性整体安装J2s和寄生阶段中的单个RNA分子。寄生虫阶段的ISH需要在感染根部提取线虫前一天采取额外的程序。我们描述了在线虫整个组织中使用地高辛(DIG)标记的cDNA探针检测转录物。

Adapting the Smart-seq2 Protocol for Robust Single Worm RNA-seq
Author:
Date:
2018-02-20
[Abstract]  Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the transcriptome of an individual worm. While developed for individual Steinernema carpocapsae and Caenorhabditis elegans larvae as well as embryos, the protocol should be adaptable for other nematode species and small invertebrates. In addition, we describe how to analyze the RNA-seq results using the Galaxy online environment. We expect that this method will be useful for the studying gene expression variances of individual nematodes in wild type and mutant backgrounds. [摘要]  大多数线虫是小蠕虫,缺乏足够的RNA用于常规的RNA-seq协议,而没有汇集成千上万的个体。 我们已经调整了Smart-seq2协议来排序单个蠕虫的转录组。 虽然针对Steinernema carpocapsae和Caenorhabditis elegans幼虫以及胚胎开发,但该方案应该适用于其他线虫物种和小无脊椎动物。 另外,我们介绍如何使用Galaxy在线环境分析RNA-seq结果。 我们预计这种方法将有助于研究野生型和突变体背景个体线虫的基因表达差异。

【背景】低输入RNA-seq方案和扩增试剂盒,例如Smart-seq(Takara Bio,USA,Inc)和SuperAmp(Miltenyl Biotec,Inc),已经越来越多地开发和商业化,作为对低输入RNA-基于小组织,单一微生物和单细胞的seq研究。这些研究经常探索并解决特定群体(例如细胞群体,复杂组织或微生物群体)的个体中的异源基因表达。针对微生物(如线虫)的低输入RNA-seq方案的改进和适应将通过允许在单一线虫水平上分析基因表达异质性而极大地有益于线虫领域。在这里,我们已经调整了单细胞RNA-seq方案Smart-seq2(Picelli等人,2013和2014; Trombetta等人,2014),对于单线虫RNA测序。我们成功地在昆虫寄生线虫Steinernema ...

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