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- W4310367057 abstract "RNA is an easily accessible molecule within the cell and therefore a commonly assayed parameter of gene expression. The classical methods of molecular biology all measure RNA abundance directly through the use of DNA and RNA probes, though there is an upper limit to the sensitivity that each of these methods can offer. This ceiling is due to the labile nature of RNA and the fact that, while variable in abundance, individual RNA molecules cannot be amplified directly in order to boost the signal. Fortunately, there is a straightforward solution to these sensitivity impediments. First, RNA can be converted enzymatically into complementary DNA (cDNA). Second, cDNA can be amplified by PCR or cloned for various types of analysis. The synthesis of cDNA is a central component of research involving molecular biology techniques. cDNA is a product of enzymatic in vitro synthesis, one strand at a time, using RNA as template material. The type of enzyme required for the synthesis of first-strand cDNA is known as an “RNA-dependent DNA polymerase”; it is far more commonly known as reverse transcriptase. That cDNA is much more stable than the corresponding RNA template material from which it is synthesized facilitates extensive analysis in both the short- and long-term. When you make cDNA, what you are really doing is making a permanent biochemical record of the cell. One makes a biochemical “snapshot” since the only cDNA that can be synthesized comes from the RNA of transcriptionally active loci. Unlike intrinsically labile RNA, cDNA can be archived for years in the freezer and when constructed with appropriately characterized ends, the cDNA can be propagated indefinitely. Transcriptionally silent loci are not represented in a cDNA pool. This chapter addresses the subtleties of making cDNA, while the next chapter addresses the subtleties of amplifying it." @default.
- W4310367057 created "2022-12-09" @default.
- W4310367057 creator A5003710396 @default.
- W4310367057 date "2023-01-01" @default.
- W4310367057 modified "2023-09-27" @default.
- W4310367057 title "cDNA: a permanent biochemical record of the cell" @default.
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- W4310367057 doi "https://doi.org/10.1016/b978-0-323-90221-2.00032-1" @default.
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