Related Experiment Video
Updated: Jul 15, 2026
19:57
An Affordable HIV-1 Drug Resistance Monitoring Method for Resource Limited Settings
Published on: March 30, 2014
DNA sequence analysis of Prinker-modified restriction fragments after collection from capillary electrophoresis with
Journal of Chromatography. A
|September 13, 1996
Summary
This study presents a novel method for DNA sequencing using capillary electrophoresis (CE) and automated fraction collection. The technique successfully sequences Prinker-modified DNA fragments, offering a new approach for genomic and mutation analysis.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- DNA sequencing is crucial for genetic research and diagnostics.
- Capillary electrophoresis (CE) offers high-resolution separation of DNA fragments.
- Automated fraction collection enhances the efficiency of isolating specific DNA molecules.
Purpose of the Study:
- To demonstrate a procedure for sequencing DNA restriction fragments isolated by automated CE.
- To validate the use of Prinker-modified fragments for subsequent sequencing.
- To establish a generalizable method for sequencing unknown or mutated DNA.
Main Methods:
- DNA restriction digestion and Prinker ligation to create cohesive ends.
- Separation of modified fragments using capillary electrophoresis (CE).
- Automated fraction collection and reinjection for validation.
- Polymerase chain reaction (PCR) amplification and cycle sequencing of isolated fragments.
Main Results:
- Successful isolation and collection of Prinker-modified DNA fragments via CE.
- High-accuracy sequencing (99.3%) of an 808 bp fragment, covering 427 and 450 bases on both strands.
- Demonstration of the feasibility of sequencing unknown genomic DNA and mutation mixtures.
Conclusions:
- Automated CE fraction collection combined with Prinker technology provides an effective method for DNA sequencing.
- This approach is applicable to sequencing diverse DNA samples, including genomic and mutated DNA.
- The developed procedure enhances the capabilities for genetic analysis and discovery.
More Related Videos
Related Concept Videos
01:24DNA 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...
03:00Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
01:05Maxam-Gilbert Sequencing
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...