Pulsed field gel electrophoresis on single murine hemopoietic colonies.
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...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
1993
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| _version_ | 1797065447430946816 |
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| author | Boultwood, J Thompson, M Fidler, C Lorimore, SA Lewis, MS Wainscoat, J Wright, E |
| author_facet | Boultwood, J Thompson, M Fidler, C Lorimore, SA Lewis, MS Wainscoat, J Wright, E |
| author_sort | Boultwood, J |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-06T21:28:47Z |
| format | Journal article |
| id | oxford-uuid:43f6e49d-6eaf-4548-a5c0-0f772f7efc62 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:28:47Z |
| publishDate | 1993 |
| record_format | dspace |
| spelling | oxford-uuid:43f6e49d-6eaf-4548-a5c0-0f772f7efc622022-03-26T14:58:51ZPulsed field gel electrophoresis on single murine hemopoietic colonies.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:43f6e49d-6eaf-4548-a5c0-0f772f7efc62EnglishSymplectic Elements at Oxford1993Boultwood, JThompson, MFidler, CLorimore, SALewis, MSWainscoat, JWright, EWe 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. |
| spellingShingle | Boultwood, J Thompson, M Fidler, C Lorimore, SA Lewis, MS Wainscoat, J Wright, E Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title | Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title_full | Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title_fullStr | Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title_full_unstemmed | Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title_short | Pulsed field gel electrophoresis on single murine hemopoietic colonies. |
| title_sort | pulsed field gel electrophoresis on single murine hemopoietic colonies |
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