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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...
Labeling DNA Probes03:31

Labeling DNA Probes

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.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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...

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Related Experiment Video

Updated: Jun 8, 2026

Combining QD-FRET and Microfluidics to Monitor DNA Nanocomplex Self-Assembly in Real-Time
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Published on: August 26, 2009

A reliable method for detecting complexed DNA in vitro.

C Holladay1, M Keeney, B Newland

  • 1Network of Excellence for Functional Biomaterials, National University of Ireland, NFB building, IDA business park, Dangan, Newcastle, Galway, Ireland.

Nanoscale
|September 14, 2010
PubMed
Summary
This summary is machine-generated.

A new method using Cy5 fluorescent labeling allows accurate quantification of both bound and unbound DNA in gene delivery systems. This overcomes limitations of traditional PicoGreen assays, enabling better characterization of biomaterial-based gene delivery.

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Area of Science:

  • Biomaterials Science
  • Gene Delivery Systems
  • Analytical Chemistry

Background:

  • Quantifying eluted nucleic acids is crucial for evaluating gene delivery systems.
  • Traditional methods like PicoGreen assays struggle with complexed DNA, hindering accurate assessment.

Purpose of the Study:

  • To develop and validate a novel method for quantifying both bound and unbound DNA in gene delivery systems.
  • To compare the efficacy of Cy5 fluorescent labeling against traditional PicoGreen assays.

Main Methods:

  • DNA was permanently labeled with the fluorescent dye Cy5 before complexation.
  • Elution of six different DNA complexes from a collagen scaffold was quantified using both Cy5 labeling and PicoGreen assays.

Main Results:

  • PicoGreen assay detected only three of six DNA complexes.
  • Cy5 fluorescent labeling successfully detected all six DNA complexes, regardless of their complexation state.
  • This enabled reliable quantification of all eluted complexes from the scaffold.

Conclusions:

  • Cy5 fluorescent labeling offers a reliable method for quantifying DNA elution, independent of DNA complexation state.
  • This technique provides a significant advancement over intercalating dyes for characterizing gene delivery systems.