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- W2912226369 abstract "The exquisite 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. The phenotype of cells is ultimately governed by the modulation of gene expression; 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 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 while favoring efficient forward hybridization kinetics, both of which are dependent upon salt concentration, hybridization temperature, hybridization time, 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. Regardless of the method utilized, there are terms and concepts common to all approaches that must be clearly understood. 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 a 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. These methods are contemporary examples of whole-transcriptome approaches." @default.
- W2912226369 created "2019-02-21" @default.
- W2912226369 creator A5003710396 @default.
- W2912226369 date "2017-01-01" @default.
- W2912226369 modified "2023-09-25" @default.
- W2912226369 title "Nonarray Methods for Global Analysis of Gene Expression" @default.
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- W2912226369 doi "https://doi.org/10.1016/b978-0-12-804678-4.00020-8" @default.
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