Related Experiment Video
Updated: Oct 25, 2025
07:10
Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
Published on: July 8, 2025
627
Recent developments in the characterization of nucleic acid hybridization kinetics
Yuan-I Chen1, Manasa P Sripati1, Trung Duc Nguyen1
1Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX 78712, USA.
Current Opinion in Biomedical Engineering
|August 9, 2021
Summary
Measuring nucleic acid (NA) hybridization kinetics in live cells is now possible using advanced single-molecule tracking. This breakthrough enables deeper understanding of cellular processes and improved biotechnologies.
Area of Science:
- Molecular Biology
- Biotechnology
Background:
- Nucleic acid (NA) hybridization is crucial for cellular functions, biotechnology, and therapeutics.
- Current methods for measuring NA hybridization kinetics are limited to ensemble measurements or non-physiological conditions.
Purpose of the Study:
- To review single-molecule approaches for measuring NA hybridization kinetics within live mammalian cells.
- To discuss the implications of these measurements for understanding cellular processes and advancing biotechnologies.
Main Methods:
- Utilizing 3D single-molecule tracking techniques to observe multiple annealing and melting events of single oligonucleotides.
- Tracking freely diffusing oligonucleotides within live mammalian cells.
Main Results:
- Recent advances enable observation of NA hybridization dynamics at the single-molecule level in real-time within live cells.
- These methods overcome limitations of ensemble measurements and non-native conditions.
Conclusions:
- Live-cell single-molecule hybridization kinetics measurements offer new opportunities for biological discovery.
- Further research is needed to address unresolved challenges and fully leverage this technology for improved biotechnologies and therapeutics.
Related Concept Videos
02:57Southern Blot
21.0K
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...
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...
21.0K
02:57Real Time RT-PCR
62.5K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...
62.5K
02:31In-situ Hybridization
9.8K
In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
9.8K