Methods for quantitation of gene expression英文文献.pdfVIP

[Frontiers in Bioscience 14, 552-569, January 1, 2009] Methods for quantitation of gene expression Vigdis Nygaard, Eivind Hovig Department of Tumor Biology, Institute for Cancer Research, Norwegian Radium Hospital, 0310 Oslo, Norway TABLE OF CONTENTS 1. Abstract 2. Measuring gene products at the transcriptional level 3. A reductionist approach versus a high throughput approach 4. Common methods for measuring mRNA levels 4.1. Subtractive hybridization 4.2. Differential display 4.3. RNase protection assay 4.4. Quantitative real time RT-PCR 4.5. Real competitive PCR 4.6. Serial analysis of gene expression (SAGE) 4.7. Massive parallel signature sequencing (MPSS) 4.8. Microarray technology 4.8.1. Principle of microarray technology and array platforms 4.8.2. Target preparation and hybridization 4.9. General technology comparison 5. Increasing technology applicability by resolving restrictions and limitations 5.1. Global mRNA amplification 6. Concluding remarks and future perspective 7. References 1. ABSTRACT 2. MEASURING GENE PRODUCTS AT THE TRANSCRIPTIONAL LEVEL Gene expression of protein encoding genes can be quantitatively measured at the transcriptional level by a Nucleic acids in the form of DNA and RNA number of low- to high-throughput methods. The control heredity and cellular function. DNA and RNA can sensitivity of each method is dependent on both the be studied in a qualitative manner, such as by investigating intrinsic properties of the