Rapid method for construction of yeast artificial chromosome human DNA libraries involving the trapping of cells in agarose films

Tatsuo Kai; Jac chan Song; Hiroyuki Tashiro; Fujiko Saka; Kazuo Ozawa; Tokiko Miwa; Issay Kitabayashi; Ei ichi Soeda; Kazushige Yokoyama

doi:10.1016/0014-5793(90)81443-R

Rapid method for construction of yeast artificial chromosome human DNA libraries involving the trapping of cells in agarose films

Tatsuo Kai, Jac chan Song, Hiroyuki Tashiro, Fujiko Saka, Kazuo Ozawa, Tokiko Miwa, Issay Kitabayashi, Ei ichi Soeda, Kazushige Yokoyama

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

5 Citations (Scopus)

Abstract

A simple method for molecular cloning of fragments of more than one hundred kilobase pairs of exogenous DNA, by the encapsulation of cell in agarose beads, was reported previously for the construction of a human genomic DNA library in a yeast artificial chromosome (YAC) vector in situ YAC construction [1]. The efficiency of this procedure is impaired by the step in which agarose beads that contain human DNA fragments are melted before transformation. The incomplete solubility of the ligated human DNA fragment. YAC vector often results in lower than desirable frequencies of transformation. To overcome this problem we have developed a new improved method that involves use of an agarose film. The technical manipulations involved in the construction of clones of very large segments of human DNA are discussed.

Original language	English
Pages (from-to)	77-82
Number of pages	6
Journal	FEBS Letters
Volume	275
Issue number	1-2
DOIs	https://doi.org/10.1016/0014-5793(90)81443-R
Publication status	Published - 26-11-1990
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(90)81443-R

Cite this

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title = "Rapid method for construction of yeast artificial chromosome human DNA libraries involving the trapping of cells in agarose films",

abstract = "A simple method for molecular cloning of fragments of more than one hundred kilobase pairs of exogenous DNA, by the encapsulation of cell in agarose beads, was reported previously for the construction of a human genomic DNA library in a yeast artificial chromosome (YAC) vector in situ YAC construction [1]. The efficiency of this procedure is impaired by the step in which agarose beads that contain human DNA fragments are melted before transformation. The incomplete solubility of the ligated human DNA fragment. YAC vector often results in lower than desirable frequencies of transformation. To overcome this problem we have developed a new improved method that involves use of an agarose film. The technical manipulations involved in the construction of clones of very large segments of human DNA are discussed.",

author = "Tatsuo Kai and Song, {Jac chan} and Hiroyuki Tashiro and Fujiko Saka and Kazuo Ozawa and Tokiko Miwa and Issay Kitabayashi and Soeda, {Ei ichi} and Kazushige Yokoyama",

year = "1990",

month = nov,

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doi = "10.1016/0014-5793(90)81443-R",

language = "English",

volume = "275",

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journal = "FEBS Letters",

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publisher = "John Wiley and Sons Inc.",

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T1 - Rapid method for construction of yeast artificial chromosome human DNA libraries involving the trapping of cells in agarose films

AU - Kai, Tatsuo

AU - Song, Jac chan

AU - Tashiro, Hiroyuki

AU - Saka, Fujiko

AU - Ozawa, Kazuo

AU - Miwa, Tokiko

AU - Kitabayashi, Issay

AU - Soeda, Ei ichi

AU - Yokoyama, Kazushige

PY - 1990/11/26

Y1 - 1990/11/26

N2 - A simple method for molecular cloning of fragments of more than one hundred kilobase pairs of exogenous DNA, by the encapsulation of cell in agarose beads, was reported previously for the construction of a human genomic DNA library in a yeast artificial chromosome (YAC) vector in situ YAC construction [1]. The efficiency of this procedure is impaired by the step in which agarose beads that contain human DNA fragments are melted before transformation. The incomplete solubility of the ligated human DNA fragment. YAC vector often results in lower than desirable frequencies of transformation. To overcome this problem we have developed a new improved method that involves use of an agarose film. The technical manipulations involved in the construction of clones of very large segments of human DNA are discussed.

AB - A simple method for molecular cloning of fragments of more than one hundred kilobase pairs of exogenous DNA, by the encapsulation of cell in agarose beads, was reported previously for the construction of a human genomic DNA library in a yeast artificial chromosome (YAC) vector in situ YAC construction [1]. The efficiency of this procedure is impaired by the step in which agarose beads that contain human DNA fragments are melted before transformation. The incomplete solubility of the ligated human DNA fragment. YAC vector often results in lower than desirable frequencies of transformation. To overcome this problem we have developed a new improved method that involves use of an agarose film. The technical manipulations involved in the construction of clones of very large segments of human DNA are discussed.

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Rapid method for construction of yeast artificial chromosome human DNA libraries involving the trapping of cells in agarose films

Abstract

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