Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q
| ISSN: |
0003-2697
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|---|---|
| Keywords: |
DNA ; FPLC ; HPLC ; Nucleic acid chemistry ; chromatography ; nucleic acids ; techniques
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| Source: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
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| Topics: |
Biology
Chemistry and Pharmacology
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| Type of Medium: |
Electronic Resource
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| URL: |
| _version_ | 1798292440165122050 |
|---|---|
| autor | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| autorsonst | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| book_url | http://dx.doi.org/10.1016/0003-2697(87)90558-6 |
| datenlieferant | nat_lic_papers |
| fussnote | Separation of DNA restriction fragments by FPLC ion-exchange chromatography on Mono Q and Mono P columns was investigated. The columns were found to be particularly suitable for the separation of fragments up to 500-600 bp long. Larger fragments can also be separated although less effectively. We found the following practical working ranges for the parameters investigated: pH, 4 to 11; flow rate, 0.05 to 0.6 ml/min corresponding to separation times between 2 and 20 h. (better resolution is achieved at lower flow rates); gradient slope; between 0.5 mm eluting salt/ml buffer and over 5 mm/ml (better resolution is achieved at lower gradient slopes; eluting ionic strength was found to be independent of gradient slope); gradient composition, chloride salts of smaller monovalent cations eluted the DNA at lower ionic strengths but separations obtained were similar; additives, substances such as urea, formamide, and EDTA can be added without chromatographic effects; sample amount: amounts from 2.5 to 200 μg were applied, corresponding to single peak content of from 42 ng to 74μg DNA. Yields were generally over 90% and the chromatographed DNA was fully accessible to restriction enzyme cleavage. Separations occurred predominantly according to DNA size, but AT-rich fragments were retarded in a predictable way. |
| hauptsatz | hsatz_simple |
| identnr | NLZ183989295 |
| issn | 0003-2697 |
| journal_name | Analytical Biochemistry |
| materialart | 1 |
| package_name | Elsevier |
| publikationsort | Amsterdam |
| publisher | Elsevier |
| reference | 166 (1987), S. 158-171 |
| schlagwort | DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques |
| search_space | articles |
| shingle_author_1 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| shingle_author_2 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| shingle_author_3 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| shingle_author_4 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. |
| shingle_catch_all_1 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques 0003-2697 00032697 Elsevier |
| shingle_catch_all_2 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques 0003-2697 00032697 Elsevier |
| shingle_catch_all_3 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques 0003-2697 00032697 Elsevier |
| shingle_catch_all_4 | Westman, E. Eriksson, S. Laas, T. Pernemalm, P.-A. Skold, S.-E. Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques DNA FPLC HPLC Nucleic acid chemistry chromatography nucleic acids nucleic acids techniques 0003-2697 00032697 Elsevier |
| shingle_title_1 | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q |
| shingle_title_2 | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q |
| shingle_title_3 | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q |
| shingle_title_4 | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q |
| sigel_instance_filter | dkfz geomar wilbert ipn albert fhp |
| source_archive | Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
| timestamp | 2024-05-06T08:48:33.406Z |
| titel | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q |
| titel_suche | Separation of DNA restriction fragments by ion-exchange chromatography on FPLC columns Mono P and Mono Q Separation of DNA restriction fragments by FPLC ion-exchange chromatography on Mono Q and Mono P columns was investigated. The columns were found to be particularly suitable for the separation of fragments up to 500-600 bp long. Larger fragments can also be separated although less effectively. We found the following practical working ranges for the parameters investigated: pH, 4 to 11; flow rate, 0.05 to 0.6 ml/min corresponding to separation times between 2 and 20 h. (better resolution is achieved at lower flow rates); gradient slope; between 0.5 mm eluting salt/ml buffer and over 5 mm/ml (better resolution is achieved at lower gradient slopes; eluting ionic strength was found to be independent of gradient slope); gradient composition, chloride salts of smaller monovalent cations eluted the DNA at lower ionic strengths but separations obtained were similar; additives, substances such as urea, formamide, and EDTA can be added without chromatographic effects; sample amount: amounts from 2.5 to 200 μg were applied, corresponding to single peak content of from 42 ng to 74μg DNA. Yields were generally over 90% and the chromatographed DNA was fully accessible to restriction enzyme cleavage. Separations occurred predominantly according to DNA size, but AT-rich fragments were retarded in a predictable way. |
| topic | W V |
| uid | nat_lic_papers_NLZ183989295 |