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Related Concept Videos

The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
Chromosome Replication02:31

Chromosome Replication

Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins.  This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin of...
Replication in Eukaryotes01:29

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...
Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview
Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview

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Related Experiment Video

Updated: Jun 22, 2026

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome
06:40

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

Published on: March 22, 2018

Replication initiation point mapping: approach and implications.

Sapna Das-Bradoo1, Anja-Katrin Bielinsky

  • 1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 1, 2009
PubMed
Summary
This summary is machine-generated.

Replication initiation point (RIP) mapping precisely identifies DNA synthesis start sites. This method, using lambda-exonuclease to purify nascent DNA, reveals where DNA replication begins in budding yeast.

Related Experiment Videos

Last Updated: Jun 22, 2026

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome
06:40

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

Published on: March 22, 2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Eukaryotic chromosome duplication initiates at multiple replication origins.
  • DNA synthesis proceeds bidirectionally from these origins.
  • Precise initiation sites within origins remain largely uncharacterized.

Purpose of the Study:

  • To present a detailed protocol for Replication Initiation Point (RIP) mapping in budding yeast.
  • To enable nucleotide-level mapping of DNA synthesis start sites.
  • To investigate the precise locations of DNA replication initiation.

Main Methods:

  • Replication Initiation Point (RIP) mapping.
  • Purification of nascent DNA initiated by RNA primers.
  • Digestion of broken DNA fragments using lambda-exonuclease, preserving nascent strands with RNA primers.

Main Results:

  • RIP mapping allows for precise nucleotide-level determination of DNA replication start sites.
  • The technique has been successfully applied across various organisms, including yeast and human cells.
  • Studies suggest the origin recognition complex (ORC) binding site is adjacent to replication start points in yeast.

Conclusions:

  • RIP mapping is a sensitive and effective technique for identifying replication origins.
  • The protocol detailed here facilitates precise mapping of DNA replication initiation in budding yeast.
  • Findings support the role of ORC in directing replication initiation to specific sites.