In vitro Biomineralization Assay
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Author:
Date:
2014-02-05
[Abstract] Biomineralization in vertebrates has both physiological and pathological aspects. Physiological mineralization is essential for proper development and function of hard tissues, such as bone, teeth, and growth plate cartilage, but it does not occur in soft tissues. Pathological ectopic mineralization, in contrast, occurs in soft tissues, including blood vessels, kidney, articular cartilage, and cardiovascular tissue. Here, we describe the simple method for detecting and measuring the presence of mineralized nodules in cardiac ventricular fibroblasts by using von Kossa and alizarin red S staining, and a colorimetric method for calcium quantification, respectively.
[摘要] 脊椎动物中的生物矿化具有生理和病理学方面。 生理矿化对于硬组织(例如骨,牙齿和生长板软骨)的正常发育和功能是必需的,但它不在软组织中发生。 相反,病理异位矿化发生在软组织中,包括血管,肾,关节软骨和心血管组织。 在这里,我们描述了使用von Kossa和茜素红S染色和钙定量的比色法分别检测和测量心室成纤维细胞中矿化结节的存在的简单方法。
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High Resolution Detection of Genetic Changes Associated with Transposons
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Author:
Date:
2013-06-05
[Abstract] Transposable elements (TEs) are repetitive sequences, capable of inducing genetic mutations through their transpositional activity, or by non-homologous or illegitimate recombination. Because of their similarity and often high copy numbers, examining the effects of mutations caused by TEs in different samples (tissues, individuals, species, etc.) can be difficult. Thus, high throughput methods have been developed for genotyping TEs in un-sequenced genomes. A common method is termed Transposon Display (or transposon SSAP), which utilizes restriction enzymes and PCR amplification to produce chimeric DNA molecules that include genomic and TE DNA. The advent of second generation sequencing technologies, such as 454-pyrosequencing, have dramatically improved the resolution of this assay, ...
[摘要] 转座元件(TE)是重复序列,能够通过其转座活性或通过非同源或非同源重组诱导遗传突变。 由于它们的相似性和通常高的拷贝数,检查由不同样品(组织,个体,物种等)中TE引起的突变的影响可能是困难的。 因此,已经开发了用于在未测序的基因组中对TE进行基因分型的高通量方法。 常见的方法称为转座子展示(或转座子SSAP),其利用限制酶和PCR扩增来产生包括基因组和TE DNA的嵌合DNA分子。 第二代测序技术(例如454焦磷酸测序)的出现已显着改善了该测定的分辨率,允许同时测序所有PCR产物,代表特定基因组中的所有扩增的TE位点。
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