In Situ Amplification of Rearranged Gene Sequences in Individual Cells for Tracing Clonal Cells in Smears from Blood and Bone Marrow
The polymerase chain reaction (PCR) has proven a valuable tool to amplify minute amounts of specific DNA to quantities which can be visualized in gels after ethidium bromide staining  and even can be analyzed by sequence analysis. Thus gene sequences altered by rearrangement or translocation typical for certain clonal expansions such as the bcr/abl translocation in chronic myeloic leukemia (CML) (t9; 22) [2,3,4] or the bcl2 rearrangement in follicular lymphoma (t14/1S)  can be traced even if only a small fraction of cells belongs to the clone in a heterogeneous population. To date the DNA is extracted from the bulk of cells for analysis and thus the altered gene sequences can not be assigned to distinct cells. However, neither in CML nor in follicular lymphoma clonal cells can for certain be distinguished from their normal counterparts by morphology and it is not known which cells carry the rearranged genome and which do not carry it.
KeywordsLymphoma Leukemia Agarose Bromide Electrophoresis
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