Abstract
DNA adducts occur through environmental, therapeutic, dietary, oxygen stress, and aging processes. A modified thin-layer chromatographic (TLC) technique can assess base composition and adduct formation. This requires labeling DNA by "shot-gun" 5'-phosphorylation of representative 32P-α-deoxyribonucleotide monophosphates. Subsequent 3'-monophosphate digest "sister exchanges" a radioactive 32PO42- to the neighboring cold nucleotide. Separation in two-dimensional polyethyleneimine-cellulose TLC is carried out in acetic acid, (NH4)2SO4, and (NH4)HSO4. The technique was applied to control DNA, cold substition of dUMP, methylation, depurination, and pBR322. This technique quantifies low-molecular-mass adducts and DNA integrity both in vivo and in vitro.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 41-55 |
| Number of pages | 15 |
| Journal | Journal of Chromatography B: Biomedical Sciences and Applications |
| Volume | 574 |
| Issue number | 1 |
| DOIs | |
| State | Published - Feb 7 1992 |
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ASJC Scopus subject areas
- Chemistry(all)
- Clinical Biochemistry
- Molecular Medicine
Cite this
Enzymatic shot-gun 5'-phosphorylation and 3'-sister phosphate exchange : a two-dimensional thin-layer chromatographic technique to measure DNA deoxynucleotide modification. / Steinberg, Jacob J.; Cajigas, Antonio; Brownlee, Michael.
In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 574, No. 1, 07.02.1992, p. 41-55.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Enzymatic shot-gun 5'-phosphorylation and 3'-sister phosphate exchange
T2 - a two-dimensional thin-layer chromatographic technique to measure DNA deoxynucleotide modification
AU - Steinberg, Jacob J.
AU - Cajigas, Antonio
AU - Brownlee, Michael
PY - 1992/2/7
Y1 - 1992/2/7
N2 - DNA adducts occur through environmental, therapeutic, dietary, oxygen stress, and aging processes. A modified thin-layer chromatographic (TLC) technique can assess base composition and adduct formation. This requires labeling DNA by "shot-gun" 5'-phosphorylation of representative 32P-α-deoxyribonucleotide monophosphates. Subsequent 3'-monophosphate digest "sister exchanges" a radioactive 32PO42- to the neighboring cold nucleotide. Separation in two-dimensional polyethyleneimine-cellulose TLC is carried out in acetic acid, (NH4)2SO4, and (NH4)HSO4. The technique was applied to control DNA, cold substition of dUMP, methylation, depurination, and pBR322. This technique quantifies low-molecular-mass adducts and DNA integrity both in vivo and in vitro.
AB - DNA adducts occur through environmental, therapeutic, dietary, oxygen stress, and aging processes. A modified thin-layer chromatographic (TLC) technique can assess base composition and adduct formation. This requires labeling DNA by "shot-gun" 5'-phosphorylation of representative 32P-α-deoxyribonucleotide monophosphates. Subsequent 3'-monophosphate digest "sister exchanges" a radioactive 32PO42- to the neighboring cold nucleotide. Separation in two-dimensional polyethyleneimine-cellulose TLC is carried out in acetic acid, (NH4)2SO4, and (NH4)HSO4. The technique was applied to control DNA, cold substition of dUMP, methylation, depurination, and pBR322. This technique quantifies low-molecular-mass adducts and DNA integrity both in vivo and in vitro.
UR - http://www.scopus.com/inward/record.url?scp=0026548505&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026548505&partnerID=8YFLogxK
U2 - 10.1016/0378-4347(92)80096-9
DO - 10.1016/0378-4347(92)80096-9
M3 - Article
C2 - 1629287
AN - SCOPUS:0026548505
VL - 574
SP - 41
EP - 55
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
SN - 0378-4347
IS - 1
ER -