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Title Guidelines for molecular analysis in archive tissues / Giorgio Stanta, editor
Published Heidelberg ; New York : Springer, 2011
Online access available from:
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Description 1 online resource (xvi, 323 pages) : illustrations (some color)
Series Issues in agroecology, Present status and future prospectus ; v. 1
Issues in agroecology. Present status and future prospectus ; v. 1
Contents Archive Tissues -- Preanalytical Time Interval (PATI) and Fixation -- Formalin-Free Fixatives -- Manual Microdissection -- Tissue Microarray (TMA) -- Laser Capture Microdissection (LCM) -- DNA Extraction from Formalin-Fixed Paraffin-Embedded (FFPE) Tissues -- DNA Extraction from Formalin-Fixed Paraffin-Embedded Tissues (FFPE) (from Small Fragments of Tissues or Microdissected Cells) -- Fast Protocol for DNA Extraction from Formalin-Fixed Paraffin-Embedded Tissues -- DNA Extraction from Blood and Forensic Samples -- Restoration and Reconstruction of DNA Length -- RNA Extraction from Formalin-Fixed Paraffin-Embedded Tissues -- RNA Extraction from Decalcified and Non-decalcified Formalin-Fixed Paraffin-Embedded Tissues -- RNA Temperature Demodification -- MicroRNA Extraction from Formalin-Fixed Paraffin-Embedded Tissues -- Quantification of Nucleic Acids -- Integrity Assessment of Nucleic Acids -- DNase Treatment of RNA -- Reverse Transcription -- General Protocol for End-Point PCR -- Quantitative End-Point PCR -- Nested-PCR -- General Protocol for Dot-Blot -- PCR-ELISA -- Quantitative Real-Time RT-PCR -- DNA Sequencing from PCR Products -- Pyrosequencing Analysis for Detection of KRAS Mutation -- Microsatellite Instability (MSI) Detection in DNA from FFPE Tissues -- General Protocol for Loss of Heterozygosity Detection -- Qualitative Methylation Status Assessment -- Quantitative Methylation Status Assessment in DNA from FFPE Tissues with Bisulfite Modification and Real-Time Quantitative MSP -- Comparative Genomic Hybridisation (CGH) -- Multiplex Ligation-dependent Probe Amplification (MLPA) -- Fluorescence In Situ Hybridization (FISH) on Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Sections -- Introduction on Genome-wide Expression Profiling from Formalin-Fixed Paraffin-Embedded Tissues Using Microarrays -- Introduction -- Protein Extraction from Formalin-Fixed Paraffin-Embedded Tissues -- Rapid Protein Extraction from Formalin-Fixed Paraffin-Embedded Tissue Samples -- Protein Extraction Protocol from Tissues Treated with Formalin-Free Fixatives -- Trichloroacetic Acid (TCA) Precipitation of Proteins -- One-Dimensional Sodium-Dodecyl-Sulfate (SDS) Polyacrylamide Gel Electrophoresis -- Visualization of Proteins -- Western Blotting -- Dot Blot -- Membrane Stripping -- Reverse-Phase Protein Microarrays -- Introduction -- Sample Preparation and In Gel Tryptic Digestion for Mass Spectrometry Experiments -- MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissues -- Introduction -- Internal Quality Control (IQC) Organization -- Standards for Equipment and Reagents
Summary A huge amount of fixed and paraffin-embedded tissue is stored in every hospital. This is very precious material that can be used for translational research and for diagnostics. The molecular methods employed for analysis of these tissues are similar to the usual molecular biology and proteomics methods, but reliable results can be obtained only if specific steps are followed with great care. ยก This book provides detailed and precise guidelines for molecular analysis of archive tissues and will serve as an invaluable aid for researchers and pathologists involved in translational research and diagnostics. The topics addressed include pre-analytical treatment of tissues, isolation of tissue components, DNA and RNA methods, proteomics methods, internal quality control procedures, and storage and treatment of reagents. Clear notes and explanations are included to simplify use of the protocols for the less experienced. The authors are a group of acknowledged experts who have developed the described methods and validated them within the European project "Archive Tissues: Improving Molecular Medicine Research and Clinical Practice - IMPACTS", which has involved 21 leading institutions in 11 countries
Bibliography Includes bibliographical references and index
Subject Pathology, Molecular.
Molecular Diagnostic Techniques.
Sequence Analysis.
Tissue Extracts.
Tissue Fixation.
Form Electronic book
Author Stanta, Giorgio.
LC no. 2011925943
ISBN 9783642178900