By John Swansbury
A suite of key cytogenetic and FISH strategies utilized by glossy scientific laboratories within the genetic research of human malignancies. The book's sensible recommendation and strategies are compatible to be used at each point of craftsmanship, together with absolutely validated laboratories, yet with a sympathetic bias in the direction of someone contemplating constructing a brand new cytogenetics provider. the following the reader will locate not just effortless tutorials at the basics of human karyotypes and chromosome research, but additionally unique discussions on how laboratories could optimally improve their repertoire of features to incorporate such more moderen complementary thoughts as CGH, FISH, and M-FISH.
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Extra resources for Cancer Cytogenetics: Methods and Protocols (Methods in Molecular Biology)
41. Langabeer, S. , Gale, R. , et al. (1997b) Frequency of CBFbeta/MYH11 fusion transcripts in patients entered into the U. K. MRC AML trials. Br. J. Haematol. 96, 738–739. 42. , Cotterill, S. , Ross, F. , et al. (2000) Cytogenetically cryptic AML1/ETO and CBFB/MYH11 gene rearrangements: incidence in 412 cases of acute myeloid leukemia. Br. J. Haematol. 111, 1051–1056. 43. , Powles, R. , et al. (1999) Outcome of biphenotypic acute leukemia. Hematologica 84, 699–706. 44. , et al. (1996) Cytogenetic findings in acute biphenotypic leukemia.
The appropriate speed and duration of centrifugation depends on the type of centrifuge that the laboratory uses; the longer the rotor arm, the greater is the centripetal force, so the speed or time can be reduced. It is important to ensure that sufficient time is given for the white cells to be collected, as they generally take longer than the red cells. , step 5, so setting the tubes to centrifuge for a few minutes longer after this step can be worthwhile. In the author’s laboratory, centrifugation takes 11 min at 12,000 rpm; the most effective speed and time would need to be determined for other centrifuges.
Certain reagents, termed mitogens, are regularly used in cytogenetics studies to stimulate lymphocytes into division, and some of these are described in Chapter 9. However, the disease may affect lymphoid cells so that they are not capable of responding to mitogens, or the treatment may suppress the immune response; in these cases mitogens will not be effective in producing divisions. If the lymphocytes have already been transformed, for example, because the patient has an infection, then lymphocyte divisions can be found in unstimulated cultures.