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- W4310367121 abstract "The orchestration of gene expression at the transcriptional level and beyond is responsible for the differentiation of cells and tissues and, ultimately, for conferring the phenotype of the organism. It is logical to focus on the mRNA population of a cell and how the complexity and composition of that population changes in response to the experimental challenge, compared to control or baseline conditions. Two types of methods are used to understand the biochemical changes that occur within a cell, namely, subtractive methods (suppression subtractive hybridization) and nonsubtractive methods (mRNA differential display). The overall efficiency of subtractive methods depends on maintaining high stringency conditions and the presence of excess driver throughout, whereas the strategy of nonsubtractive methods for gene expression profiling is more about selective amplification rather than subtraction. The tester is the cDNA population containing induced sequences that are to be sequestered. The driver is the control or reference population of cDNA sequences against which the tester is being compared. During the subtraction hybridization, the driver is always present in large molar excess over the tester. This will ensure first-order hybridization kinetics, resulting in complete hybridization and concomitant removal of all sequences common to both populations. In the case of mRNA differential display, multiple PCR reactions with different combinations of anchored first-strand cDNA primers generate different subsets of products that are easily resolved by polyacrylamide gel electrophoresis for a side-by-side comparison of two or more samples using fluorescent-tagged primers or through the incorporation of radiolabel. These new gene discovery techniques are contemporary examples of whole-transcriptome approaches that do not require prior knowledge of the genes that will be investigated." @default.
- W4310367121 created "2022-12-09" @default.
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- W4310367121 date "2023-01-01" @default.
- W4310367121 modified "2023-10-18" @default.
- W4310367121 title "Subtractive and nonsubtractive methods for the analysis of gene expression" @default.
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- W4310367121 doi "https://doi.org/10.1016/b978-0-323-90221-2.00033-3" @default.
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