RT-PCR Protocols
Preface
Contents
Contributors
INTRODUCTION
1 RT-PCR in Biomedicine Opportunities Arising from the New Accessibility of mRNA
2 The Basics of RT-PCR
HIGHLY SENSITIVE DETECTION AND ANALYSIS OF MRNA
3 Using the Quantitative Competitive RT-PCR Technique to Analyze Minute Amounts of Different mRNAs in Small Tissue Samples
4 Detection of mRNA Expression and Alternative Splicing in a Single Cell
5 Nested RT-PCR Sensitivity Controls are Essential to Determine the Biological Significance of Detected mRNA
QUANTITATIVE RT-PCR
6 Quantitative RT-PCR
7 Rapid Development of a Quantitative-Competitive (qc) RT-PCR Assay Using a Composite Primer Approach
8 Quantitation of Gene Expression by RT-PCR and HPLC Analysis of PCR Products
9 Time-Resolved Fluorometric Detection of Cytokine mRNAs Amplified by RT-PCR
10 Mimic-Based RT-PCR Quantitation of Substance P mRNA in Human Mononuclear Phagocytes and Lymphocytes
DETECTION AND ANALYSIS OF RNA VIRUSES
11 Detection and Quantification of the Hepatitis C Viral Genome
12 Semi-Quantitative Detection of Hepatitis C Virus RNA by βReal-Timeβ RT-PCR
13 RT-PCR for the Assessment of Genetically Heterogenous Populations of the Hepatitis C Virus
IN SITU LOCALIZATION OF MRNA EXPRESSION
14 In Situ Immuno-PCR A Newly Developed Method for Highly Sensitive Antigen Detection In Situ
15 RT-PCR from Laser-Capture Microdissected Samples
16 Mycobacterium paratuberculosis Detected by Nested PCR in Intestinal Granulomas Isolated by LCM in Cases of Crohnβs Disease
17 RT-PCR-Based Approaches to Generate Probes for mRNA Detection by In Situ Hybridization
DIFFERENTIAL MRNA EXPRESSION
18 Amplified RNA for Gene Array Hybridizations
19 Semi-Quantitative Determination of Differential Gene Expression in Primary Tumors and Matched Metastases by RT-PCR Comparison with Other Methods
GENETIC ANALYSIS
20 Detection of Single Nucleotide Polymorphisms Using a Non-Isotopic RNase Cleavage Assay
RT-PCR IN IMMUNOLOGY
21 Detection of Clonally Expanded T-Cells by RT-PCR-SSCP and Nucleotide Sequencing of T-Cell Receptor β’-CDR3 Regions
22 Generation of scFv from a Phage Display Mini-Library Derived from Tumor-Infiltrating B-Cells
23 Generation of Murine scFv Intrabodies from B-Cell Hybridomas
24 Quantitation of mRNA Levels by RT-PCR in Cells Purified by FACS Application to Peripheral Cannabinoid Receptors in Leukocyte Subsets
RT-PCR IN ANTI-SENSE TECHNOLOGY
25 Detection of Anti-Sense RNA Transcripts by Anti-Sense RT-PCR
RT-PCR IN CDNA CLONING
26 RT-PCR in cDNA Library Construction
27 An RT-PCR-Based Protocol for the Rapid Generation of Large, Representative cDNA Libraries for Expression Screening
Index