Molecular stretching of long DNA in agarose gel using alternating current electric fields

Noritada Kaji, Masanori Ueda, Yoshinobu Baba

Research output: Contribution to journalArticle

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Abstract

We demonstrate a novel method for stretching a long DNA molecule in agarose gel with alternating current (AC) electric fields. The molecular motion of a long DNA (T4 DNA; 165.6 kb) in agarose gel was studied using fluorescence microscopy. The effects of a wide range of field frequencies, field strengths, and gel concentrations were investigated. Stretching was only observed in the AC field when a frequency of -10 Hz was used. The maximal length of the stretched DNA had the longest value when a field strength of 200 to 400 V/cm was used. Stretching was not sensitive to a range of agarose gel concentrations from 0.5 to 3%. Together, these experiments indicate that the optimal conditions for stretching long DNA in an AC electric field are a frequency of 10 Hz with a field strength of 200 V/cm and a gel concentration of 1% agarose. Using these conditions, we were able to successfully stretch Saccharomyces cerevisiae chromosomal DNA molecules (225-2,200 kb). These results may aid in the development of a novel method to stretch much longer DNA, such as human chromosomal DNA, and may contribute to the analysis of a single chromosomal DNA from a single cell.

Original languageEnglish
Pages (from-to)335-344
Number of pages10
JournalBiophysical Journal
Volume82
Issue number1
DOIs
Publication statusPublished - Jan 1 2002

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Sepharose
Gels
DNA
Fluorescence Microscopy
Saccharomyces cerevisiae

All Science Journal Classification (ASJC) codes

  • Biophysics

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Molecular stretching of long DNA in agarose gel using alternating current electric fields. / Kaji, Noritada; Ueda, Masanori; Baba, Yoshinobu.

In: Biophysical Journal, Vol. 82, No. 1, 01.01.2002, p. 335-344.

Research output: Contribution to journalArticle

Kaji, Noritada ; Ueda, Masanori ; Baba, Yoshinobu. / Molecular stretching of long DNA in agarose gel using alternating current electric fields. In: Biophysical Journal. 2002 ; Vol. 82, No. 1. pp. 335-344.
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