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Related Concept Videos

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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...
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...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...

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Updated: Jun 18, 2026

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction
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Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

Published on: November 20, 2018

In vitro double transposition for DNA identification.

Nicholas J Heredia1, N Reginald Beer, Christine A Hara

  • 1Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.

Analytical Biochemistry
|December 1, 2009
PubMed
Summary
This summary is machine-generated.

A novel double transposition technique enables DNA sequencing by creating priming sites within the target DNA. This in vitro method simplifies the process, reduces hands-on time, and maintains genome integrity for efficient analysis.

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Last Updated: Jun 18, 2026

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Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach
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Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Published on: September 11, 2017

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Current DNA sequencing methods can be time-consuming and require complex procedures.
  • Maintaining the integrity of the target genome is crucial for accurate sequencing.
  • In vitro methods offer potential for streamlined and automated biological assays.

Purpose of the Study:

  • To develop an efficient in vitro method for DNA sequencing using transposition.
  • To create a technique that amplifies target DNA regions between two transposons.
  • To demonstrate the application of this method in microfluidic platforms.

Main Methods:

  • A double transposition technique was employed to insert two distinct transposons into target DNA.
  • The transposition reaction, DNA repair, and sequencing were performed entirely in vitro.
  • The method was adapted for high-throughput applications using microfluidic droplet technology.

Main Results:

  • The double transposition technique successfully generated priming sites for amplifying the region between transposons.
  • The target DNA genome remained intact throughout the in vitro process.
  • The method significantly reduced assay time and the number of enzymatic steps compared to standard techniques.
  • Transposition reactions were demonstrated in picoliter droplets on a microfluidic platform.

Conclusions:

  • The double transposition technique provides an efficient and streamlined in vitro approach for DNA sequencing.
  • This method preserves genome integrity and is amenable to automation and high-throughput applications.
  • The in vitro transposition strategy offers a faster and simpler alternative to existing sequencing preparation methods.