Chromatin: A Practical ApproachHannah Gould Chromatin comprises the complex of nucleic acids and proteins that make up the chromosomes, and regulation of its structure is increasingly recognized as an important factor in gene expression. Researchers from related fields needing an introduction to the methodology of chromatin will findthis book invaluable, as will experienced workers. Contents: Isolation and fractionation of chromatin and linker histones; Extraction and separation of core histones and non-histone chromosomal proteins; Mapping chromatin structure in Drosophila; In vivo footprint and chromatin analysis by LMPCR; Analysis of nuclear scaffold attachment regions;Electron microscopy of chromatin; Counting nucleosome cores on circular DNA using topoisomerase I; In vitro reconstitution of chromatin and analysis of nucleosome positioning; Transcriptional and structural analysis of chromatin assembled in vivo; An assembly of chromatin and nuclear structures inXenopus egg extracts; Appendix: List of Suppliers; Index |
Contents
In vivo footprint and chromatin analysis | 4 |
Isolation and sizefractionation of chromatin | 11 |
Removal of linker histones from native chromatin | 20 |
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acetate Acids Res acrylamide agarose gel aliquots analysis binding Biochem Biol cells centrifugation chicken erythrocyte chloroquine chromatin chromatin assembly chromatin structure cleavage column concentration containing core histones core-particle described in Protocol dialysis digestion dilution DNA fragments DNase Drosophila EDTA elution EMBO end-labelled Equipment and reagents ethanol ethidium bromide Figure filter footprinting fractions gene gradient grids histone H1 histone octamer homogenizer hybridization Incubate isolation linker histones LMPCR membrane method mg/ml MgCl2 microcentrifuge micrococcal nuclease MNase molecules monomer NaCl Nucl nuclear nuclei nucleosome nucleosome cores oligonucleotide phosphorylated plasmid PMSF polymerase prepared primer extension probe procedure proteins purified reaction reconstituted chromatin region Remove replication restriction enzyme Resuspend the pellet room temperature rotor samples scaffolds Sigma Sorvall sperm spermatid step stock solution sucrose supercoils supernatant TBE buffer template tion topoisomerase topoisomers transcription Tris-HCl tube volume Xenopus µg/ml