Rapid isolation of highly pure single-stranded DNA from phagemids

Bisheng Zhou; Qiaoxiang Dong; Ruihua Ma; Yuanhong Chen; Junhua Yang; Lu Zhe Sun; Changjiang Huang

doi:10.1016/j.ab.2009.03.044

Rapid isolation of highly pure single-stranded DNA from phagemids

Bisheng Zhou, Qiaoxiang Dong, Ruihua Ma, Yuanhong Chen, Junhua Yang, Lu Zhe Sun, Changjiang Huang

Department of Cell Systems & Anatomy

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

8 Scopus citations

Abstract

Single-stranded DNA (ssDNA) has many applications in molecular biology and biotechnology. The conventional method for the preparation of ssDNA from phagemids is laborious, costly, and inefficient. Here we describe an integrated protocol for consistent production of phagemid ssDNA from a bacteria/phagemid/help phage complex and rapid isolation and purification of the ssDNA with a silica column followed by duplex-specific nuclease digestion. The major advantages of our method are the expediency, low cost, and consistent yield of highly pure ssDNA that is suitable for direct sequencing and other applications. This method is especially useful for large-scale preparation of high-quality ssDNA.

Original language	English (US)
Pages (from-to)	177-179
Number of pages	3
Journal	Analytical Biochemistry
Volume	389
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2009.03.044
State	Published - Jun 15 2009

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2009.03.044

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title = "Rapid isolation of highly pure single-stranded DNA from phagemids",

abstract = "Single-stranded DNA (ssDNA) has many applications in molecular biology and biotechnology. The conventional method for the preparation of ssDNA from phagemids is laborious, costly, and inefficient. Here we describe an integrated protocol for consistent production of phagemid ssDNA from a bacteria/phagemid/help phage complex and rapid isolation and purification of the ssDNA with a silica column followed by duplex-specific nuclease digestion. The major advantages of our method are the expediency, low cost, and consistent yield of highly pure ssDNA that is suitable for direct sequencing and other applications. This method is especially useful for large-scale preparation of high-quality ssDNA.",

author = "Bisheng Zhou and Qiaoxiang Dong and Ruihua Ma and Yuanhong Chen and Junhua Yang and Sun, {Lu Zhe} and Changjiang Huang",

note = "Funding Information: This work was supported by the major project of the Science and Technology Department of Zhejiang Province (2008C03001-2), key project of the Natural Science Foundation of Zhejiang Province (Z2080266), key project of the Science and Technology Department of Wenzhou (S20060023), 5010 key project of Wenzhou Medical College (XNK06009), and a National Institutes of Health (NIH) grant (R01CA079683).",

year = "2009",

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doi = "10.1016/j.ab.2009.03.044",

language = "English (US)",

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journal = "Analytical Biochemistry",

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T1 - Rapid isolation of highly pure single-stranded DNA from phagemids

AU - Zhou, Bisheng

AU - Dong, Qiaoxiang

AU - Ma, Ruihua

AU - Chen, Yuanhong

AU - Yang, Junhua

AU - Sun, Lu Zhe

AU - Huang, Changjiang

N1 - Funding Information: This work was supported by the major project of the Science and Technology Department of Zhejiang Province (2008C03001-2), key project of the Natural Science Foundation of Zhejiang Province (Z2080266), key project of the Science and Technology Department of Wenzhou (S20060023), 5010 key project of Wenzhou Medical College (XNK06009), and a National Institutes of Health (NIH) grant (R01CA079683).

PY - 2009/6/15

Y1 - 2009/6/15

N2 - Single-stranded DNA (ssDNA) has many applications in molecular biology and biotechnology. The conventional method for the preparation of ssDNA from phagemids is laborious, costly, and inefficient. Here we describe an integrated protocol for consistent production of phagemid ssDNA from a bacteria/phagemid/help phage complex and rapid isolation and purification of the ssDNA with a silica column followed by duplex-specific nuclease digestion. The major advantages of our method are the expediency, low cost, and consistent yield of highly pure ssDNA that is suitable for direct sequencing and other applications. This method is especially useful for large-scale preparation of high-quality ssDNA.

AB - Single-stranded DNA (ssDNA) has many applications in molecular biology and biotechnology. The conventional method for the preparation of ssDNA from phagemids is laborious, costly, and inefficient. Here we describe an integrated protocol for consistent production of phagemid ssDNA from a bacteria/phagemid/help phage complex and rapid isolation and purification of the ssDNA with a silica column followed by duplex-specific nuclease digestion. The major advantages of our method are the expediency, low cost, and consistent yield of highly pure ssDNA that is suitable for direct sequencing and other applications. This method is especially useful for large-scale preparation of high-quality ssDNA.

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Rapid isolation of highly pure single-stranded DNA from phagemids

Abstract

ASJC Scopus subject areas

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