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Related Concept Videos

Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...

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Single Molecule Analysis of Laser Localized Psoralen Adducts
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On-chip laser-induced DNA dehybridization.

E K Wheeler1, B R Baker, W T Piggott

  • 1Center for Micro and Nano Technology, LLNL, Livermore, California, USA.

The Analyst
|May 22, 2013
PubMed
Summary
This summary is machine-generated.

Researchers developed a focused Infrared (IR) laser method to selectively recover hybridized DNA from specific spots on DNA microarrays. This technique enables targeted DNA sequencing and reduces errors in downstream analysis.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • DNA microarrays detect pathogens and genetic markers via nucleic acid hybridization.
  • Simultaneous detection of multiple targets is common in molecular diagnostics.
  • Emergent organisms can yield novel hybridization patterns requiring further analysis.

Purpose of the Study:

  • To develop a method for localized DNA recovery from DNA microarrays.
  • To enable targeted sequencing of specific hybridization events.
  • To reduce background noise and improve downstream sequencing accuracy.

Main Methods:

  • Utilized a focused Infrared (IR) laser to selectively heat spots on a DNA microarray.
  • Induced localized dehybridization of bound DNA at user-selected locations.
  • Recovered the dehybridized DNA into an elution buffer for subsequent analysis.

Main Results:

  • Successfully demonstrated localized DNA dehybridization and recovery using a focused IR laser.
  • Minimized disruption to adjacent spots on the microarray.
  • Prepared recovered DNA for downstream amplification and sequencing.

Conclusions:

  • Focused IR laser dehybridization is a novel technique for targeted DNA recovery from microarrays.
  • This method enhances the analysis of unknown or emergent genetic targets.
  • The technique shows potential for application in protein microarrays as well.