Ricicová Markéta, Palková Zdena
Department of Genetics and Microbiology, Charles University, Vinicná 5, 12844 Prague 2, Czech Republic.
FEMS Yeast Res. 2003 Oct;4(1):119-22. doi: 10.1016/S1567-1356(03)00145-4.
Precise quantification and quality characterisation of isolated RNAs are prerequisites for their further exploitation in genome-wide microarrays, Northern blots, cDNA library preparation and others. Our data indicate that RNA analyses using Agilent RNA Nano Assay exhibit several advantages when compared with those performed on ethidium bromide-stained agarose gel electrophoresis or on a spectrophotometer. The RNA Nano Assay makes it possible to estimate RNA concentrations in the range from 1000 ng microl(-1) to 17 ng microl(-1). The presence of impurities including traces of DNA within RNA samples does not influence the concentration measurements. Like agarose gel electrophoresis, RNA Nano Assay allows to analyse RNAs dissolved in formamide and therefore protected against RNase action. Moreover, it allows a clearer distinction of partially degraded samples. The limitation of RNA Nano Assay is the impossibility to detect and to analyse double-stranded RNAs.
对分离出的RNA进行精确量化和质量表征，是其在全基因组微阵列、Northern印迹、cDNA文库制备等方面进一步应用的前提条件。我们的数据表明，与在溴化乙锭染色的琼脂糖凝胶电泳或分光光度计上进行的RNA分析相比，使用安捷伦RNA纳米检测法进行RNA分析具有几个优点。RNA纳米检测法能够估算出浓度范围在1000 ng/μl至17 ng/μl之间的RNA浓度。RNA样品中包括微量DNA在内的杂质的存在，不会影响浓度测量。与琼脂糖凝胶电泳一样，RNA纳米检测法能够分析溶解在甲酰胺中因而受到核糖核酸酶作用保护的RNA。此外，它能更清晰地区分部分降解的样品。RNA纳米检测法的局限性在于无法检测和分析双链RNA。
