Rapid preparation of covalently closed circular DNA by acridine yellow affinity chromatography

Walter S. Vincent; Elliott S. Goldstein

doi:10.1016/0003-2697(81)90121-4

Rapid preparation of covalently closed circular DNA by acridine yellow affinity chromatography

Walter S. Vincent
, Elliott S. Goldstein

Life Sciences, School of (SOLS)

Research output: Contribution to journal › Article › peer-review

32 Scopus citations

Abstract

Covalently closed circular DNA can be isolated rapidly from cell lysates in a two-step process. Hydroxylapatite chromatography to prepurify the plasmid DNA from contaminating protein and RNA is followed by a step gradient elution of covalently closed circular (CCC) plasmid DNA from an acridine yellow affinity column. This procedure results in CCC DNA of a purity comparable to that obtained from ethidium bromide-CsCl gradients without lengthy centrifugation and free of contaimination by intercalating dye. Up to 250 μg of CCC pBR 322 can be isolated from 500 ml of bacterial culture in 4-6 h.

Original language	English (US)
Pages (from-to)	123-127
Number of pages	5
Journal	Analytical Biochemistry
Volume	110
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(81)90121-4
State	Published - Jan 1 1981

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Rapid preparation of covalently closed circular DNA by acridine yellow affinity chromatography",

abstract = "Covalently closed circular DNA can be isolated rapidly from cell lysates in a two-step process. Hydroxylapatite chromatography to prepurify the plasmid DNA from contaminating protein and RNA is followed by a step gradient elution of covalently closed circular (CCC) plasmid DNA from an acridine yellow affinity column. This procedure results in CCC DNA of a purity comparable to that obtained from ethidium bromide-CsCl gradients without lengthy centrifugation and free of contaimination by intercalating dye. Up to 250 μg of CCC pBR 322 can be isolated from 500 ml of bacterial culture in 4-6 h.",

author = "Vincent, \{Walter S.\} and Goldstein, \{Elliott S.\}",

note = "Funding Information: This investigation was supported by a grant from the Graduate College and by NIH Grant HD 09157.",

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doi = "10.1016/0003-2697(81)90121-4",

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AU - Vincent, Walter S.

AU - Goldstein, Elliott S.

N1 - Funding Information: This investigation was supported by a grant from the Graduate College and by NIH Grant HD 09157.

PY - 1981/1/1

Y1 - 1981/1/1

N2 - Covalently closed circular DNA can be isolated rapidly from cell lysates in a two-step process. Hydroxylapatite chromatography to prepurify the plasmid DNA from contaminating protein and RNA is followed by a step gradient elution of covalently closed circular (CCC) plasmid DNA from an acridine yellow affinity column. This procedure results in CCC DNA of a purity comparable to that obtained from ethidium bromide-CsCl gradients without lengthy centrifugation and free of contaimination by intercalating dye. Up to 250 μg of CCC pBR 322 can be isolated from 500 ml of bacterial culture in 4-6 h.

AB - Covalently closed circular DNA can be isolated rapidly from cell lysates in a two-step process. Hydroxylapatite chromatography to prepurify the plasmid DNA from contaminating protein and RNA is followed by a step gradient elution of covalently closed circular (CCC) plasmid DNA from an acridine yellow affinity column. This procedure results in CCC DNA of a purity comparable to that obtained from ethidium bromide-CsCl gradients without lengthy centrifugation and free of contaimination by intercalating dye. Up to 250 μg of CCC pBR 322 can be isolated from 500 ml of bacterial culture in 4-6 h.

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ER -

Rapid preparation of covalently closed circular DNA by acridine yellow affinity chromatography

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