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We report a technique which allows for the direct molecular analysis of single whole murine hemopoietic colonies by pulsed field gel electrophoresis (PFGE). Murine bone marrow cells were plated out in semi-solid agarose and gave rise to macroscopic colonies after 11 days in culture. Single colonies were excised from the agarose using a sterile blade and embedded without further manipulation in molten low-melting-temperature agarose. The leucocyte DNA contained within the agarose plug was subjected to restriction enzyme digestion and PFGE. Sufficient high molecular weight DNA is afforded by this method to achieve a hybridization signal with a single copy probe. This method will make PFGE directly applicable to the clonal analysis of chromosomal aberrations in hemopoietic stem and progenitor cells.

Type

Journal article

Journal

Leukemia

Publication Date

10/1993

Volume

7

Pages

1635 - 1636

Keywords

Animals, Bone Marrow, Bone Marrow Cells, Colony-Forming Units Assay, DNA, Electrophoresis, Gel, Pulsed-Field, Hematopoietic Stem Cells, Mice, Sepharose