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

Noritada Kaji, Masanori Ueda, Yoshinobu Baba

研究成果: Contribution to journalArticle査読

32 被引用数 (Scopus)

抄録

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.

本文言語英語
ページ(範囲)335-344
ページ数10
ジャーナルBiophysical Journal
82
1
DOI
出版ステータス出版済み - 2002
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生物理学

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