Density transfer as a method to analyze the progression of DNA replication forks.
The density transfer technique is a valuable tool to examine the progression of individual DNA replication forks. It is based on the transfer of cells from a medium containing dense isotopes to a medium with light (normal) isotopes (or vice versa), to obtain DNA sequences hybrid in density that can be identified as replicated molecules. Using specific DNA probes along a chromosome, the dense isotope transfer method allows determining the extent of replication at any position of a replicon and the rate of replication fork progression. In the eukaryotic model budding yeast, this technique has been useful to establish a role for different proteins during the elongation of chromosomal replication and to analyze the movement and stability of DNA replication forks under different experimental conditions.
Tercero
José Antonio
JA
Centro de Biología Molecular Severo Ochoa, CSIC/UAM, Cantoblanco, Madrid, Spain.
D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient; D015825:Chromosomes, Fungal / Q000378:metabolism; D003470:Culture Media; D015342:DNA Probes; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D009587:Nitrogen Isotopes; D009693:Nucleic Acid Hybridization; D012093:Replicon; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
D002247:Carbon Isotopes; D003470:Culture Media; D015342:DNA Probes; D004271:DNA, Fungal; D009587:Nitrogen Isotopes; D029701:Saccharomyces cerevisiae Proteins