Recovery of intact yeast chromosomal DNA from agarose gel plugs using coil-globule transition

Jun Komatsu, Michihiko Nakano, Hirofumi Kurita, Kazunori Takashima, Shinji Katsura, Akira Mizuno

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In the current studies, we designed a new approach for sizing and isolating chromosomal DNA using coil-globule transition, which avoids fragmentation of giant DNA due to mechanical stress. Although coil-globule transition is reversible and globular DNA is tolerant to mechanical stress, globular DNA cannot be manipulated by an electric field because of the loss of its negative charges. In our system, however, DNA is extracted from an agarose gel in a coiled state into a solution of PEG, and coil-globule transition is induced by cations generated at the anode. This method achieves buffer exchange without stirring, which is the main cause of mechanical stress. Real-time analysis of T4dc viral DNA molecules revealed that they change immediately from a coiled to a globular form when the cation concentration is sufficiently high. This method was used to prepare yeast chromosomal DNA in a globular state without fragmentation.

Original languageEnglish
Pages (from-to)4296-4303
Number of pages8
JournalElectrophoresis
Volume26
Issue number22
DOIs
Publication statusPublished - Nov 2005
Externally publishedYes

Fingerprint

Yeast
Sepharose
Yeasts
Gels
Mechanical Stress
Recovery
DNA
Cations
Viral DNA
DNA Fragmentation
Buffers
Electrodes
Polyethylene glycols
Anodes
Electric fields
Molecules

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Recovery of intact yeast chromosomal DNA from agarose gel plugs using coil-globule transition. / Komatsu, Jun; Nakano, Michihiko; Kurita, Hirofumi; Takashima, Kazunori; Katsura, Shinji; Mizuno, Akira.

In: Electrophoresis, Vol. 26, No. 22, 11.2005, p. 4296-4303.

Research output: Contribution to journalArticle

Komatsu, Jun ; Nakano, Michihiko ; Kurita, Hirofumi ; Takashima, Kazunori ; Katsura, Shinji ; Mizuno, Akira. / Recovery of intact yeast chromosomal DNA from agarose gel plugs using coil-globule transition. In: Electrophoresis. 2005 ; Vol. 26, No. 22. pp. 4296-4303.
@article{e71d403588c341719c08dedd4ea4cb42,
title = "Recovery of intact yeast chromosomal DNA from agarose gel plugs using coil-globule transition",
abstract = "In the current studies, we designed a new approach for sizing and isolating chromosomal DNA using coil-globule transition, which avoids fragmentation of giant DNA due to mechanical stress. Although coil-globule transition is reversible and globular DNA is tolerant to mechanical stress, globular DNA cannot be manipulated by an electric field because of the loss of its negative charges. In our system, however, DNA is extracted from an agarose gel in a coiled state into a solution of PEG, and coil-globule transition is induced by cations generated at the anode. This method achieves buffer exchange without stirring, which is the main cause of mechanical stress. Real-time analysis of T4dc viral DNA molecules revealed that they change immediately from a coiled to a globular form when the cation concentration is sufficiently high. This method was used to prepare yeast chromosomal DNA in a globular state without fragmentation.",
author = "Jun Komatsu and Michihiko Nakano and Hirofumi Kurita and Kazunori Takashima and Shinji Katsura and Akira Mizuno",
year = "2005",
month = "11",
doi = "10.1002/elps.200500061",
language = "English",
volume = "26",
pages = "4296--4303",
journal = "Electrophoresis",
issn = "0173-0835",
publisher = "Wiley-VCH Verlag",
number = "22",

}

TY - JOUR

T1 - Recovery of intact yeast chromosomal DNA from agarose gel plugs using coil-globule transition

AU - Komatsu, Jun

AU - Nakano, Michihiko

AU - Kurita, Hirofumi

AU - Takashima, Kazunori

AU - Katsura, Shinji

AU - Mizuno, Akira

PY - 2005/11

Y1 - 2005/11

N2 - In the current studies, we designed a new approach for sizing and isolating chromosomal DNA using coil-globule transition, which avoids fragmentation of giant DNA due to mechanical stress. Although coil-globule transition is reversible and globular DNA is tolerant to mechanical stress, globular DNA cannot be manipulated by an electric field because of the loss of its negative charges. In our system, however, DNA is extracted from an agarose gel in a coiled state into a solution of PEG, and coil-globule transition is induced by cations generated at the anode. This method achieves buffer exchange without stirring, which is the main cause of mechanical stress. Real-time analysis of T4dc viral DNA molecules revealed that they change immediately from a coiled to a globular form when the cation concentration is sufficiently high. This method was used to prepare yeast chromosomal DNA in a globular state without fragmentation.

AB - In the current studies, we designed a new approach for sizing and isolating chromosomal DNA using coil-globule transition, which avoids fragmentation of giant DNA due to mechanical stress. Although coil-globule transition is reversible and globular DNA is tolerant to mechanical stress, globular DNA cannot be manipulated by an electric field because of the loss of its negative charges. In our system, however, DNA is extracted from an agarose gel in a coiled state into a solution of PEG, and coil-globule transition is induced by cations generated at the anode. This method achieves buffer exchange without stirring, which is the main cause of mechanical stress. Real-time analysis of T4dc viral DNA molecules revealed that they change immediately from a coiled to a globular form when the cation concentration is sufficiently high. This method was used to prepare yeast chromosomal DNA in a globular state without fragmentation.

UR - http://www.scopus.com/inward/record.url?scp=28244486013&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28244486013&partnerID=8YFLogxK

U2 - 10.1002/elps.200500061

DO - 10.1002/elps.200500061

M3 - Article

VL - 26

SP - 4296

EP - 4303

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 22

ER -