Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems.
In this paper, a two-step process for initial capture of plasmid DNA (pDNA) and partial removal of RNA using polyethylene glycol (PEG) and di-potassium hydrogen phosphate aqueous two-phase systems (ATPS) has been investigated. A Kühni-type ATPS extraction column was prepared with 50 ml (12% (w/w) PE...
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| Format: | Article |
| Language: | English English |
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Elsevier BV
2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/17505/ http://psasir.upm.edu.my/id/eprint/17505/1/Two.pdf |
| _version_ | 1848843259965603840 |
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| author | Luechau, Frank Ling, Tau Chuan Lyddiatt, Andrew |
| author_facet | Luechau, Frank Ling, Tau Chuan Lyddiatt, Andrew |
| author_sort | Luechau, Frank |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | In this paper, a two-step process for initial capture of plasmid DNA (pDNA) and partial removal of RNA using polyethylene glycol (PEG) and di-potassium hydrogen phosphate aqueous two-phase systems (ATPS) has been investigated. A Kühni-type ATPS extraction column was prepared with 50 ml (12% (w/w) PEG 1450, 12% (w/w) phosphate) of stationary phase and loaded with crude mobile phase (26% (w/w) PEG 1450, 4% (w/w) phosphate and 70% (w/w) lysate) at a flow rate of 6 ml min−1 at an impeller speed of 200 rpm. The experiment was terminated after 100 min, and after complete resettling of the phases, 45 ml of stationary phase was harvested. During a subsequent second extraction step contained 18% (w/w) PEG 300 and 14% (w/w) phosphate, a proportion of RNA, which was also concentrated during the column process, was removed. It was demonstrated that the recovery of pDNA in the second bottom phase was 89.4%, which was similar to the initial recovery after column extraction (92.1%). |
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| format | Article |
| id | upm-17505 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:12:12Z |
| publishDate | 2010 |
| publisher | Elsevier BV |
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| spelling | upm-175052015-10-05T04:29:07Z http://psasir.upm.edu.my/id/eprint/17505/ Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. Luechau, Frank Ling, Tau Chuan Lyddiatt, Andrew In this paper, a two-step process for initial capture of plasmid DNA (pDNA) and partial removal of RNA using polyethylene glycol (PEG) and di-potassium hydrogen phosphate aqueous two-phase systems (ATPS) has been investigated. A Kühni-type ATPS extraction column was prepared with 50 ml (12% (w/w) PEG 1450, 12% (w/w) phosphate) of stationary phase and loaded with crude mobile phase (26% (w/w) PEG 1450, 4% (w/w) phosphate and 70% (w/w) lysate) at a flow rate of 6 ml min−1 at an impeller speed of 200 rpm. The experiment was terminated after 100 min, and after complete resettling of the phases, 45 ml of stationary phase was harvested. During a subsequent second extraction step contained 18% (w/w) PEG 300 and 14% (w/w) phosphate, a proportion of RNA, which was also concentrated during the column process, was removed. It was demonstrated that the recovery of pDNA in the second bottom phase was 89.4%, which was similar to the initial recovery after column extraction (92.1%). Elsevier BV 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/17505/1/Two.pdf Luechau, Frank and Ling, Tau Chuan and Lyddiatt, Andrew (2010) Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. Process Biochemistry, 45 (8). pp. 1432-1436. ISSN 1359-5113 10.1016/j.procbio.2010.05.015 English |
| spellingShingle | Luechau, Frank Ling, Tau Chuan Lyddiatt, Andrew Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title | Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title_full | Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title_fullStr | Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title_full_unstemmed | Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title_short | Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems. |
| title_sort | two-step process for initial capture of plasmid dna and partial removal of rna using aqueous two-phase systems. |
| url | http://psasir.upm.edu.my/id/eprint/17505/ http://psasir.upm.edu.my/id/eprint/17505/ http://psasir.upm.edu.my/id/eprint/17505/1/Two.pdf |