Double strand breaks induced by low doses of γ rays or heavy ions: Quantitation in nonradioactive human DNA

Betsy M. Sutherland; Paula V. Bennett; John C. Sutherland

doi:10.1006/abio.1996.0290

Double strand breaks induced by low doses of γ rays or heavy ions: Quantitation in nonradioactive human DNA

Betsy M. Sutherland, Paula V. Bennett, John C. Sutherland

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

17 Scopus citations

Abstract

We have developed a method of quantitating low frequencies (0-30 sites/10⁸ base pairs) of double strand breaks in ~1 μg of nonradioactive human DNA. Unirradiated or irradiated DNA is digested with the restriction endonuclease NotI, producing cleavage fragments that include a major group centered at ~1.2-1.3 Mbp. The DNA molecules are separated as a function of size by transverse alternating field electrophoresis. The frequency of double strand breaks is computed directly from the decrease in number average molecular length induced in the 1.2- to 1.3-Mbp cleavage fragment group by ¹³⁷Cs γ or Fe²⁶ (1.1 GeV/nucleon) irradiation vs the corresponding unirradiated DNA samples. The double strand break frequency can be quantitated easily in the dose range of 0-10 cGy of γ rays. The frequency of breaks per unit dose calculated for γ irradiation of DNA in human cells (~4.6 double strand breaks/10⁹ bp/Gy) is within the range of values obtained by others (2-8 sites/10⁹ bp/Gy) who used methods requiring higher doses.

Original language	English (US)
Pages (from-to)	53-60
Number of pages	8
Journal	Analytical Biochemistry
Volume	239
Issue number	1
DOIs	https://doi.org/10.1006/abio.1996.0290
State	Published - Jul 15 1996
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Double strand breaks induced by low doses of γ rays or heavy ions: Quantitation in nonradioactive human DNA",

abstract = "We have developed a method of quantitating low frequencies (0-30 sites/108 base pairs) of double strand breaks in ~1 μg of nonradioactive human DNA. Unirradiated or irradiated DNA is digested with the restriction endonuclease NotI, producing cleavage fragments that include a major group centered at ~1.2-1.3 Mbp. The DNA molecules are separated as a function of size by transverse alternating field electrophoresis. The frequency of double strand breaks is computed directly from the decrease in number average molecular length induced in the 1.2- to 1.3-Mbp cleavage fragment group by 137Cs γ or Fe26 (1.1 GeV/nucleon) irradiation vs the corresponding unirradiated DNA samples. The double strand break frequency can be quantitated easily in the dose range of 0-10 cGy of γ rays. The frequency of breaks per unit dose calculated for γ irradiation of DNA in human cells (~4.6 double strand breaks/109 bp/Gy) is within the range of values obtained by others (2-8 sites/109 bp/Gy) who used methods requiring higher doses.",

author = "Sutherland, {Betsy M.} and Bennett, {Paula V.} and Sutherland, {John C.}",

note = "Funding Information: This research was supported by the Of{\textregistered}ce of Health and Environmental Research of the U.S. Department of Energy, by a National Institutes of Health Grant HG00371 to J.C.S., and by the Space Radiation Health Program, Life and Biomedical Sciences and Applications Division of the National Aeronautics and Space Administration. We thank R. Sautkulis, Biology, BNL for irradiating samples with 137Cs g rays and providing dosimetry, Dr. Jack Miller, Lawrence Berkeley Laboratory, for providing dosimetric measurements for heavy ion irradiation, and Dr. Richard Setlow for critical reading of the manuscript.",

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AU - Sutherland, Betsy M.

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N1 - Funding Information: This research was supported by the Of®ce of Health and Environmental Research of the U.S. Department of Energy, by a National Institutes of Health Grant HG00371 to J.C.S., and by the Space Radiation Health Program, Life and Biomedical Sciences and Applications Division of the National Aeronautics and Space Administration. We thank R. Sautkulis, Biology, BNL for irradiating samples with 137Cs g rays and providing dosimetry, Dr. Jack Miller, Lawrence Berkeley Laboratory, for providing dosimetric measurements for heavy ion irradiation, and Dr. Richard Setlow for critical reading of the manuscript.

PY - 1996/7/15

Y1 - 1996/7/15

N2 - We have developed a method of quantitating low frequencies (0-30 sites/108 base pairs) of double strand breaks in ~1 μg of nonradioactive human DNA. Unirradiated or irradiated DNA is digested with the restriction endonuclease NotI, producing cleavage fragments that include a major group centered at ~1.2-1.3 Mbp. The DNA molecules are separated as a function of size by transverse alternating field electrophoresis. The frequency of double strand breaks is computed directly from the decrease in number average molecular length induced in the 1.2- to 1.3-Mbp cleavage fragment group by 137Cs γ or Fe26 (1.1 GeV/nucleon) irradiation vs the corresponding unirradiated DNA samples. The double strand break frequency can be quantitated easily in the dose range of 0-10 cGy of γ rays. The frequency of breaks per unit dose calculated for γ irradiation of DNA in human cells (~4.6 double strand breaks/109 bp/Gy) is within the range of values obtained by others (2-8 sites/109 bp/Gy) who used methods requiring higher doses.

AB - We have developed a method of quantitating low frequencies (0-30 sites/108 base pairs) of double strand breaks in ~1 μg of nonradioactive human DNA. Unirradiated or irradiated DNA is digested with the restriction endonuclease NotI, producing cleavage fragments that include a major group centered at ~1.2-1.3 Mbp. The DNA molecules are separated as a function of size by transverse alternating field electrophoresis. The frequency of double strand breaks is computed directly from the decrease in number average molecular length induced in the 1.2- to 1.3-Mbp cleavage fragment group by 137Cs γ or Fe26 (1.1 GeV/nucleon) irradiation vs the corresponding unirradiated DNA samples. The double strand break frequency can be quantitated easily in the dose range of 0-10 cGy of γ rays. The frequency of breaks per unit dose calculated for γ irradiation of DNA in human cells (~4.6 double strand breaks/109 bp/Gy) is within the range of values obtained by others (2-8 sites/109 bp/Gy) who used methods requiring higher doses.

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Double strand breaks induced by low doses of γ rays or heavy ions: Quantitation in nonradioactive human DNA

Abstract

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