TY - JOUR
T1 - Electrophoretic analyses of nucleosomes and other protein-DNA complexes
AU - Huang, S. Y.
AU - Garrard, W. T.
N1 - Funding Information:
This work was supported by Grant No. 6M38984.01 of the National Institutes of Health (H.N.) and No. 3.348-0.86 of the Swiss National Science Foundation and by the Kanton Basel (M.N.). Hans Noll is a lifetime Career Professor of the American Cancer Society.
Funding Information:
We are indebted to the following colleagues from this laboratory for important contributions in perfecting the techniques described here: Richard Todd, Stephen Albfight, Phyllis Nelson, Tim Reudelhuber, Alan Davis, Teni Boulikas, Dorothy Ball, David Gross, Stephen Rose, Peter Cockerill, Darel Hunting, Christopher Szent-Gyorgyi,C harles Lynch, and Mary Barnard. We thank Veronica Blasquez, Barbara Fishel, and Christopher Szent-Gyorgyif or review and Marie Rotondi and Susan Alexander for the preparation of the manuscript. This work has been supported by Grants GM22201, GM29935, GM25829, and GM31689 from the National Institutes of Health and Grant 1-823 from The Robert A. Welch Foundation.
PY - 1989/1/1
Y1 - 1989/1/1
N2 - Electrophoresis has proven to be a powerful resolving technique for both analytical and preparative biochemical experiments. Electrophoretic resolution of ribosome and polysome components was successfully achieved. This chapter discusses the following topics: (1) one-dimensional electrophoretic resolution of nucleosomes, (2) enzymatic preparation of chromatin and chromatin fractionation, (3) two-dimensional electrophoretic resolution of DNA and detection of specific sequences, (4) two-dimensional electrophoretic resolution of proteins using Triton-acid-urea gels, and (5) further applications for electrophoretic analyses. This chapter also have described techniques to (1) determine the overall electrophoretie patterns of various nucleoprotein species, (2) analyze the bulk DNA lengths of such species by performing a second dimension of electrophoresis in the presence of SDS, (3) determine the position of specific sequences in such two-dimensional displays after DNA transfer and nucleic acid hybridization, and (4) analyze the bulk protein compositions of such species by performing a second dimension of electrophoresis in Triton-acidurea gels.
AB - Electrophoresis has proven to be a powerful resolving technique for both analytical and preparative biochemical experiments. Electrophoretic resolution of ribosome and polysome components was successfully achieved. This chapter discusses the following topics: (1) one-dimensional electrophoretic resolution of nucleosomes, (2) enzymatic preparation of chromatin and chromatin fractionation, (3) two-dimensional electrophoretic resolution of DNA and detection of specific sequences, (4) two-dimensional electrophoretic resolution of proteins using Triton-acid-urea gels, and (5) further applications for electrophoretic analyses. This chapter also have described techniques to (1) determine the overall electrophoretie patterns of various nucleoprotein species, (2) analyze the bulk DNA lengths of such species by performing a second dimension of electrophoresis in the presence of SDS, (3) determine the position of specific sequences in such two-dimensional displays after DNA transfer and nucleic acid hybridization, and (4) analyze the bulk protein compositions of such species by performing a second dimension of electrophoresis in Triton-acidurea gels.
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U2 - 10.1016/0076-6879(89)70044-6
DO - 10.1016/0076-6879(89)70044-6
M3 - Article
C2 - 2770536
AN - SCOPUS:0024351482
SN - 0076-6879
VL - 170
SP - 116
EP - 142
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -