Molecular Analysis of CancerJacqueline Boultwood, Carrie Fidler Over the past 20 years, technological advances in molecular biology have proven invaluable to the understanding of the pathogenesis of human cancer. The application of molecular technology to the study of cancer has not only led to advances in tumor diagnosis, but has also provided markers for the assessment of prognosis and disease progression. The aim of Molecular Ana- sis of Cancer is to provide a comprehensive collection of the most up-to-date techniques for the detection of molecular changes in human cancer. Leading researchers in the field have contributed chapters detailing practical pro- dures for a wide range of state-of-the-art techniques. Molecular Analysis of Cancer includes chapters describing techniques for the identification of chromosomal abnormalities and comprising: fluor- cent in situ hybridization (FISH), spectral karyotyping (SKY), comparative genomic hybridization (CGH), and microsatellite analysis. FISH has a pro- nent role in the molecular analysis of cancer and can be used for the detection of numerical and structural chromosomal abnormalities. The recently described SKY, in which all human metaphase chromosomes are visualized in specific colors, allows for the definition of all chromosomal rearrangements and marker chromosomes in a tumor cell. Protocols for the detection of chromosomal re- rangements by PCR and RT-PCR are described, as well as the technique of DNA fingerprinting, a powerful tool for studying somatic genetic alterations in tumorigenesis. |
Contents
| 2 | |
Spectral Karyotyping in Cancer Cytogenetics | 29 |
Binaifer R Balsara Jianming Pei and Joseph R Testa 5 Detection of Chromosomal Deletions by Microsatellite Analysis | 45 |
Detection and Quantification of LeukemiaSpecific Rearrangements | 67 |
Detection of t25p23q35 Translocation by LongRange | 97 |
Use of DNA Fingerprinting to Detect Genetic Rearrangements | 107 |
Mutation Analysis of Large Genomic Regions in Tumor DNA Using | 115 |
Mutational Analysis of Oncogenes and Tumor Suppressor Genes | 125 |
Detection of Differentially Expressed Genes in Cancer Using | 179 |
Genomewide Gene Expression Analysis Using cDNA Microarrays | 195 |
Gene Expression Profiling in Cancer Using cDNA Microarrays | 205 |
Wilms Tumor Gene WT1 as a Tumor Marker for Leukemic Blast Cells | 223 |
Detection of Aberrant Methylation of the p15INK4B Gene Promoter | 239 |
Clonality Studies in Cancer Based on X Chromosome Inactivation | 251 |
Telomere Length Changes in Human Cancer | 271 |
Measurement of Telomerase Activity in Human Hematopoietic Cells | 279 |
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Common terms and phrases
acute leukemia agarose gel allele amplification approx assay band BCR-ABL bone marrow cDNA cell lines centrifuge chromosome chronic myelogenous leukemia chronic myeloid cleavage clonality cloning coverslip cytogenetic denaturing detection DGGE dH2O differential display disease distilled water DNA sequence dNTP EDTA enzyme ethanol ethidium bromide exon fluorescence fragments G6PD gel electrophoresis gene expression Genet genomic DNA human hybridization inactivation Incubate labeled leukemic marker metaphase method methylation mg/mL microarray microsatellite Molecular Analysis mRNA mutations normal Note nucleotide Ohyashiki oligonucleotide PCR products pellet peripheral blood plates polymerase chain reaction polymorphism primer probe protein Protocols quantitative reagents RNase room temperature RT-PCR samples slides sodium Southern blot SSCP stem cells supernatant TAE buffer target technique telomerase activity telomere telomere length template tion tissue transcription translocation Tris-HCl tube tumor suppressor gene wash wild-type Wilms WT1 expression levels WT1 gene µg/mL