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Related Concept Videos

Recombinant DNA01:09

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Overview
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Homologous Recombination02:31

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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...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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DNA Isolation01:24

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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...
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Labeling DNA Probes03:31

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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Southern Blot02:57

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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.
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Related Experiment Video

Updated: May 5, 2026

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
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DNA detection using recombination proteins.

Olaf Piepenburg1, Colin H Williams, Derek L Stemple

  • 1ASM Scientific Ltd, Cambridge, United Kingdom.

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Recombinase polymerase amplification (RPA) offers a simple, rapid, and instrument-free method for DNA amplification and detection. This novel technology enables sensitive nucleic acid testing outside specialized labs, advancing portable diagnostics.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostic Assays

Background:

  • Established DNA amplification techniques often require complex equipment and procedures, limiting their use in non-specialized settings.
  • The need for accessible, portable nucleic acid testing is critical for widespread disease detection and monitoring.

Purpose of the Study:

  • To introduce and validate a novel DNA amplification method, recombinase polymerase amplification (RPA).
  • To demonstrate the efficacy of RPA coupled with a probe-based detection system for instrument-free DNA testing.

Main Methods:

  • Developed recombinase polymerase amplification (RPA), combining isothermal recombinase-driven primer targeting with strand-displacement DNA synthesis.
  • Integrated RPA with a probe-based detection system for real-time monitoring.
  • Applied the combined RPA amplification/detection system to detect methicillin-resistant Staphylococcus aureus (MRSA).

Main Results:

  • RPA achieves exponential DNA amplification without sample DNA pretreatment, operating at a constant low temperature.
  • The assay demonstrated high sensitivity, detecting fewer than ten copies of genomic DNA.
  • A simple sandwich assay allowed for instrument-free detection of RPA products.

Conclusions:

  • RPA is a sensitive, specific, rapid, and isothermal DNA amplification technology.
  • The instrument-free RPA system offers a significant advancement for portable and widely accessible nucleic acid-based testing.
  • This technology holds promise for applications beyond specialized laboratories, including point-of-care diagnostics.