By Lynda J. Campbell (auth.), Lynda J. Campbell (eds.)
Cytogenetic reports of malignancy became a vital software within the medical administration of melanoma sufferers. Cancer Cytogenetics: tools and Protocols provides eminently useful key cytogenetic and FISH strategies for each degree of diagnostic provider. specialists within the box describe specified cytogenetic research tools, fluorescence in situ hybridization and array equipment at present being utilized to enquire and diagnose diversified types of melanoma. Written within the hugely winning tools in Molecular Biology™ sequence structure, chapters include introductions to their respective themes, lists of the mandatory fabrics and reagents, and step by step, with no trouble reproducible laboratory protocols. The authors of a number of the chapters have additionally supplied huge notes to steer people who are new to those equipment in the course of the pitfalls that bedevil all such testing.
Authoritative and obtainable, Cancer Cytogenetics: tools and Protocols serves as an amazing consultant to scientists of all backgrounds, letting them both determine new ideas of their laboratories or locate the various diversifications of normal tools necessary in enhancing their results.
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Additional info for Cancer Cytogenetics: Methods and Protocols
7. Cytogenetic Monitoring of CML Cytogenetic analysis of bone marrow or peripheral blood cells allows identification of the Ph translocation and other chromosome changes that are associated with the leukemic process. As the leukemic cells are naturally dividing, the addition of mitogens such as phytohemagglutinin is unnecessary, and in CML may stimulate nonleukemic populations of lymphocytes. 2) is best visualized following cell culture and when leukemic cells are arrested at metaphase, the stage of cell division when chromosomal DNA is most contracted.
This is because ABL1–BCR fusion is not possible for some variant translocations when 5¢ABL1 and 3¢BCR are on different chromosomes, when large deletions extending from within ABL1 or BCR occur, or for different conformational reasons ((8, 9, 19); see also below). Opinions are currently mixed as to whether or not the ABL1–BCR transcript, when present, is translated into a stable ABL1–BCR fusion protein (19, 20). 2. Variant Ph Translocations and Complex BCR–ABL1 Rearrangements In about 10% of cases, BCR–ABL1 fusion arises from more complex rearrangements than those that give rise to the standard t(9;22).
3. The FISH Method Always read the probe manufacturer’s protocols carefully. The method described here is robust and generally reliable, but some probes may need alterations to the procedure (see Note 2). 1. Slide Preparation For FISH studies of metaphases, standard culturing, harvesting, and fixation procedures are used, and these are described in detail in other chapters in this book. If a rapid result is needed and can be obtained from interphase cells, it is not necessary to wait to collect cells in division: simply suspend the cells in hypotonic KCl for 10 min, fix using 3:1 absolute methanol:glacial acetic acid, and then change the fixative at least three times.
Cancer Cytogenetics: Methods and Protocols by Lynda J. Campbell (auth.), Lynda J. Campbell (eds.)