Non-isotopic restriction mapping of cosmid DNA

A. Takahashi-Fujii, N. Maeda, Yoshizumi Ishino, H. Kotani, I. Kato

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

A protocol for the non-isotopic restriction mapping of cosmid DNA was developed. After digestion with λ terminase and partial digestion with restriction enzymes. DNA fragments containing right or left cohesive cos termini were selectively captured by hybridization with biotinylated oligonucleotides, bound to magnetic beads coated with streptavidin and recovered by heating. Recovered DNA fragments containing cos ends were resolved by agarose gel electrophoresis, and fluorescence from the DNA fragments in the ethidium bromide-stained gel was detected with the fluorescent image analyzer, FMBIO(TM). Restriction maps were directly determined from the ladder of the partial digestion products. Two micrograms of cosmid DNA for each partial digestion were sufficient for mapping the restriction enzyme sites. This procedure provides a prototype for other cos sequence-mediated or other specific sequence-mediated DNA isolation technologies and a convenient method for non-isotopic DNA analysis. The rapid physical analysis of cosmid DNA that we present here will contribute to large DNA structural analyses like the human genome project.

元の言語英語
ページ(範囲)910-912+914
ジャーナルBioTechniques
16
発行部数5
出版物ステータス出版済み - 1994
外部発表Yes

Fingerprint

Restriction Mapping
Cosmids
DNA
Digestion
Gels
Human Genome Project
Ethidium
Streptavidin
Agar Gel Electrophoresis
DNA sequences
Ladders
Enzymes
Electrophoresis
Oligonucleotides
Sepharose
Heating
Genes
Fluorescence
Technology

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

これを引用

Takahashi-Fujii, A., Maeda, N., Ishino, Y., Kotani, H., & Kato, I. (1994). Non-isotopic restriction mapping of cosmid DNA. BioTechniques, 16(5), 910-912+914.

Non-isotopic restriction mapping of cosmid DNA. / Takahashi-Fujii, A.; Maeda, N.; Ishino, Yoshizumi; Kotani, H.; Kato, I.

:: BioTechniques, 巻 16, 番号 5, 1994, p. 910-912+914.

研究成果: ジャーナルへの寄稿記事

Takahashi-Fujii, A, Maeda, N, Ishino, Y, Kotani, H & Kato, I 1994, 'Non-isotopic restriction mapping of cosmid DNA', BioTechniques, 巻. 16, 番号 5, pp. 910-912+914.
Takahashi-Fujii A, Maeda N, Ishino Y, Kotani H, Kato I. Non-isotopic restriction mapping of cosmid DNA. BioTechniques. 1994;16(5):910-912+914.
Takahashi-Fujii, A. ; Maeda, N. ; Ishino, Yoshizumi ; Kotani, H. ; Kato, I. / Non-isotopic restriction mapping of cosmid DNA. :: BioTechniques. 1994 ; 巻 16, 番号 5. pp. 910-912+914.
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