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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Kuschak, T I
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Isolation of extrachromosomal elements by histone immunoprecipitation
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| 264 |
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|c 2001
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| 336 |
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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|b nc
|2 rdacarrier
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|a Date Completed 04.12.2001
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a Here, we describe a gentle and effective method for the rapid and reproducible isolation of histone-bound extrachromosomal DNA molecules called extrachromosomal elements (EEs). This method facilitates the harvest of a specific population of EEs following their isolation from cultured cells, primary tissues, and tumor cells. Active EEs are bound to histone proteins, and these histone-bound EEs carry actively transcribing genes such as c-myc. Our method exploits the presence of histones on EEs and serves as a first-step purification procedure, allowing for the cloning or multivariant analysis of an immunopurified sample of EEs. We isolated EEs from 4-hydroxytamoxifen (4-HT)-activated Myc-ER-regulatable Pre-B ABM cells. Following one round of immunoprecipitation, we demonstrate the purification of histone-bound EEs. We confirmed that our purification enriched for EEs that carry genes by fluorescent in situ hybridization of EEs (FISH-EEs), and we probed non-enriched and immunopurified EEs with a dihydrofolate reductase (DHFR) cDNA probe that is known to detect extrachromosomal amplification in Myc-activated cells. We demonstrate the enrichment of immunoprecipitated DHFR-containing extrachromosomal DNA molecules
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a Histones
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|a Indoles
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|a Nerve Tissue Proteins
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|a Proto-Oncogene Proteins c-myc
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|a Kuschak, B C
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Smith, G M
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Wright, J A
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Mai, S
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t BioTechniques
|d 1993
|g 30(2001), 5 vom: 01. Mai, Seite 1064-8, 1070-2
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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| 773 |
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|g volume:30
|g year:2001
|g number:5
|g day:01
|g month:05
|g pages:1064-8, 1070-2
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