生物信息学技术

Abstract Table of Contents Materials Figures Literature Cited Abstract Solid?phase synthesis of 3??aminoalkyl, 3??thioalkyl, and 3??polyethyleneglycol are described. The first two are important as specifi

Abstract Table of Contents Figures Literature Cited Abstract Conjugation of oligonucleotides at the 3 terminus is less common because this site is used for covalent linkage to solid?phase oligonucleotide synthesis supports. However, 3?olig

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit gives protocols for the attachment of intercalating and photoreactive conjugate groups to oligodeoxyribonucleotides. Protoc

Abstract Table of Contents Figures Literature Cited Abstract Reporter and conjugate groups can be added directly to the 5? terminus of oligonucleotides by appropriate modification. Conjugate groups can be used to increase the affinity of c

Abstract Table of Contents Materials Figures Literature Cited Abstract Because it is convenient to assemble nucleosomal templates through salt dialysis, large amounts of chromatin complexes can be made ea

Abstract Table of Contents Materials Literature Cited Abstract In vitro analysis of DNA in chromatin is often important for understanding mechanisms of regulation of transcription and other processes that occur on DNA. The basic unit of ch

Abstract Table of Contents Materials Figures Literature Cited Abstract The native chromatin complex within most eukaryotic nuclei is very difficult to study by biochemical means, so researchers have devel

Abstract Table of Contents Materials Figures Literature Cited Abstract Due to their similarities in size and charge, complete resolution of histones by electrophoresis poses a considerable challenge. The

Abstract Table of Contents Materials Figures Literature Cited Abstract Genetic manipulations, including gene knock?outs and mutant screens, provide an initial hint as to the function of a gene product and

Abstract Table of Contents Materials Figures Literature Cited Abstract Phage?based expression cloning is a simple, rapid, and powerful technique to identify interacting proteins. A protein of interest is

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes the use of proteins fused to glutathione?S?transferase (GST fusion proteins) to affinity purify other proteins, a t

Abstract Table of Contents Materials Figures Literature Cited Abstract Affinity chromatography is a very effective and straightforward means of purifying a protein based on its sequence?specific DNA?bindi

Abstract Table of Contents Materials Literature Cited Abstract To detect DNA binding activity, radiolabeled protein is incubated with specific DNA fragments, and protein?DNA complexes are separated from free protein by electrophoresis in n

Abstract Table of Contents Materials Figures Literature Cited Abstract Nitrocellulose binds proteins but not double?stranded DNA. Use of radioactively labeled double?stranded DNA fragments allows quantita

Abstract Table of Contents Materials Figures Literature Cited Abstract Short fragments of DNA?either natural or formed from oligonucleotides?containing a high?affinity site for a DNA?binding protein provi

Abstract Table of Contents Materials Figures Literature Cited Abstract Irradiation of protein?nucleic acid complexes with ultraviolet light causes covalent bonds to form between the nucleic acid and prote

Abstract Table of Contents Materials Figures Literature Cited Abstract Deoxyribonuclease I (DNase I) protection mapping, or footprinting, is a valuable technique for locating the specific binding sites of

Abstract Table of Contents Materials Figures Literature Cited Abstract Interference assays identify specific residues in the DNA binding site that, when modified, interfere with binding of the protein. Th

Abstract Table of Contents Materials Figures Literature Cited Abstract The DNA?binding assay using nondenaturing polyacrylamide gel electrophoresis (PAGE) provides a simple, rapid, and extremely sensitive

Abstract Table of Contents Materials Figures Literature Cited Abstract FRET microscopy enables the detection of different biochemical states of proteins in cells. The use of fluorescence in the detection