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Sterile serological pipets

Company: Olympus
Catalog#: 12-104
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Using RNA Sequencing and Spike-in RNAs to Measure Intracellular Abundance of lncRNAs and mRNAs
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Date:
2020-10-05
[Abstract]  Long noncoding RNAs (lncRNAs) play essential roles in normal physiology and in disease but their mechanisms of action can be challenging to identify. For mechanistic studies, it is often useful to know a lncRNA’s intracellular abundance, i.e., approximately how many molecules of the lncRNA are present in a typical cell of a cell-type of interest. At least two approaches have been used to approximate lncRNA intracellular abundance: single-molecule sensitivity RNA fluorescence in situ hybridization (smFISH) and single-gene, calibrated reverse-transcription followed by quantitative PCR (RT-qPCR). However, like all experimental approaches, these methods have their limitations. smFISH, when analyzed using diffraction-limited microscopy, can underestimate intracellular ... [摘要]  [摘要]长非编码RNA(lncRNA)在正常生理和疾病中起着至关重要的作用,但其作用机理可能难以鉴定。对于机理研究,了解lncRNA的胞内丰度(即在目标细胞类型的典型细胞中大约存在多少个lncRNA分子)通常很有用。至少两种方法已用于估算lncRNA细胞内丰度:单分子敏感性RNA荧光原位杂交(smFISH ...

Isolation of Primary Human Skeletal Muscle Cells
Author:
Date:
2017-11-05
[Abstract]  Primary myoblast culture is a valuable tool in research of muscle disease, pathophysiology, and pharmacology. This protocol describes techniques for dissociation of cells from human skeletal muscle biopsies and enrichment for a highly myogenic population by fluorescence-activated cell sorting (FACS). We also describe methods for assessing myogenicity and population expansion for subsequent in vitro study. [摘要]  原代成肌细胞培养是研究肌肉疾病,病理生理学和药理学的有用工具。 该协议描述了通过荧光激活细胞分选(FACS)从人类骨骼肌活检中分离细胞并富集高度肌原细胞的技术。 我们还描述了用于评估随后的体外研究中肌原性和群体扩张的方法。
【背景】来自肌肉活组织检查的原代人成肌细胞是模拟体外人体肌肉疾病的有价值的资源。成肌细胞增殖,分化和融合的改变是许多神经肌肉疾病所共有的特征,并且可以用于测定基于细胞的和药理学的治疗。人类骨骼肌活组织检查,尤其是那些受疾病影响的人,通常含有大量的非肌原细胞,如脂肪细胞和成纤维细胞。因此,纯化肌原细胞进行骨骼肌发育和疾病的体外研究是非常重要的。肌肉疾病的早期研究涉及使用组织外植体或未纯化的分离细胞(Geiger和Garvin,1957; Herrmann等人,1960; Goyle等人,1967; Bishop 1971年),后来,Blau和Webster引入了一种预先电镀技术去除成纤维细胞(Blau and ...

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