Amplification of RNA
Amplification of RNA
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DOI：
10.1016/b978-0-12-372180-8.50007-x
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
E. Kawasaki
中科院分区：
文献类型：
--
作者：
E. Kawasaki
用于检测和分析RNA分子的灵敏方法是大多数细胞/分子生物学研究的重要方面。通常使用的方法包括原位杂交、北方凝胶、斑点或狭缝印迹、S-1核酸酶测定和RNA酶A保护研究。这些技术的详细描述可在几本实验室手册中找到（Davis等，1986; Ausubel等，1987; Berger和Kimmel 1987）。这些方法中最敏感的是原位杂交，其中可以在单个细胞中检测到10到100个分子。然而，原位杂交方法在技术上可能是困难的，并且不适合于处理大量样品。使用其他技术，检测水平为约0.1至1.0 pg的靶序列。对于平均大小的mRNA，这翻译为105至10个靶序列分子，并且对于大多数实际目的，已经达到检测极限。
Sensitive methods for the detection and analysis of RNA molecules are an important aspect of most cell/molecular biology studies. Methods commonly in use include in situ hybridization, Northern gels, dot or slot blots, S-1 nuclease assays, and RNase A protection studies. Detailed descriptions of these techniques can be found in several laboratory manuals (Davis et al. 1986; Ausubel et al. 1987; Berger and Kimmel 1987). The most sensitive of these methods is in situ hybridization, in which 10 to 100 molecules can be detected in a single cell. However, the in situ hybridization method can be technically difficult and does not lend itself to the processing of a large number of samples. With other techniques, the level of detection is about 0.1 to 1.0 pg of the target sequence. For an average-sized mRNA this translates to 105 to 10 target sequence molecules, and for most practical purposes the detection limit has been reached.