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About this book
The molecular basis of disease is the driving force in biomedical research. Gene detection, quantification, and localization are required to determine disease pathogenesis. In as much as DNA is the genetic potential in a particular organism, RNA and protein expression are the functional result of this genetic programme. This book describes methods to quantify and localize gene expression, with a particular emphasis on the sensitive detection of low abundance gene expression. Sensitive detection of gene expression has been hindered by poor recovery or preservation of target sequences. The text describes optimized methods to ensure full recovery and preservation of targets from specimen acquisition through detection of specific targets and final analysis. This text also focuses upon in situ analysis of cells and tissues
Contents
Instrumentation ** Fixation for "in situ" Molecular Analysis ** A Method for Reduction of Green Wavelength Authofluorescence Emission in Cellular Preparations ** Gene Quantification: Choosing the Target ** Detection and Quantification of cytokine-producing Cells by Immunostaining ** Kaposi's Sarcoma-Associated Human Herpes Virus Eight: Localization of Viral Gene-Expression and Viral Latency ** Pathogenetic Mechanisms of Animal Orbiviruses that Cause Disease at Low Copy Number ** Localization of Hepatitis C Virus using Reverse Transcritase Fluorescence "in situ 5" Nuclease Assay (RTFISNA) ** Simultaneous Quantification and Localization of HIV gag-pol Expression Using Ultrasensative Fluorescence in situ Hybridization ** Quantification and Localization of Gene Expression: Future Directions
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