Corporate Banner
Satellite Banner
Genomics
Scientific Community
 
Become a Member | Sign in
Home>News>This Article
  News
Return

Accurate Detection of Extremely Rare Mitochondrial DNA Deletions Associated with Aging

Published: Thursday, September 05, 2013
Last Updated: Thursday, September 05, 2013
Bookmark and Share
The study published in Aging Cell identifies a new tool to accurately analyze extremely rare mitochondrial DNA deletions associated with a range of diseases and disorders as well as aging.

This approach, which relies on Droplet Digital PCR (ddPCR™) technology, will help researchers explore mitochondrial DNA (mtDNA) deletions as potential disease biomarkers.

The accumulation of mtDNA mutations is associated with aging, neuromuscular disorders, and cancer. However, methods to probe the underlying mechanisms behind this mutagenesis have been limited by their inability to accurately quantify and characterize new deletion events, which may occur at a frequency as low as one deletion event per 100 million mitochondrial genomes in normal tissue. To address these limitations, researchers at the Seattle, Washington-based Fred Hutchinson Cancer Research Center developed a ddPCR-based assay known as "Digital Deletion Detection" (3D) that allows for the high-resolution analysis of these rare deletions.

"It is incredibly difficult to study mtDNA mutations, let alone deletions, within the genome," said Dr. Jason Bielas, Assistant Member of the Public Health Sciences Division at the Fred Hutchinson Cancer Research Center and lead author of the study. "Our 3D assay shows significant improvement in specificity, sensitivity, and accuracy over conventional methods such as those that rely on real-time PCR."

Bielas added, "The increase in throughput afforded by droplet digital PCR shortened the analysis of deletion events to days compared to months using previous digital PCR methods. Without the technology, we could not have made this discovery."

At the center of the study was Bio-Rad Laboratories' QX100™ ddPCR system. Using the QX100 system, Bielas and his team analyzed eight billion human brain mtDNA genomes and identified more than 100,000 genomes with a deletion. They discovered that, contrary to popular belief, the majority of the increase in mtDNA deletions was not caused by new deletions but rather by the expansion of previous deletions. They hypothesized that the expansion of pre-existing mutations should be considered as the primary factor contributing to age-related accumulation of mtDNA deletions.

How the 3D Assay Works
3D is a novel three-step process that includes enrichment for deletion-bearing molecules, single-molecule partitioning of genomes into droplets for direct quantification via ddPCR, and breakpoint characterization using next-generation sequencing.

Once the enrichment process is completed using methods previously developed by Bielas and colleagues, the concentration of molecules within the droplets is adjusted by using the QX100 system so that the majority of droplets contain no mutant genomes while a small fraction contain only one. This process allows each deletion to be amplified without bias and without introducing the artifacts that are common in qPCR.

Following amplification, deletions can be analyzed using ddPCR to determine the absolute concentration of mutated molecules. Using the relationship between droplet fluorescence and amplicon size, Bielas and his team were able to characterize the size and complexity (whether they were a result of a few clonal expansions or a large collection of random deletions) of rare mitochondrial deletions in human brain samples.

The 3D assay provides an important new tool that will allow researchers to better study the mechanisms of deletion formation and expansion, and their role in aging. Droplet digital PCR's high throughput and increased sensitivity will also allow Bielas' lab to target other low-level disease-causing mtDNA deletions in skeletal muscle, brain tissue, and blood.


Further Information

Join For Free

Access to this exclusive content is for Technology Networks Premium members only.

Join Technology Networks Premium for free access to:

  • Exclusive articles
  • Presentations from international conferences
  • Over 3,900+ scientific posters on ePosters
  • More than 5,300+ scientific videos on LabTube
  • 35 community eNewsletters


Sign In



Forgotten your details? Click Here
If you are not a member you can join here

*Please note: By logging into TechnologyNetworks.com you agree to accept the use of cookies. To find out more about the cookies we use and how to delete them, see our privacy policy.

Related Content

Assessing the Effectiveness of Genome-Editing Technologies
Researchers have developed a cost-effective and rapid method for assessing edits generated by CRISPR-Cas9 and other genome-editing technologies.
Thursday, June 30, 2016
Awards for Bio-Rad's Protein Expression Products and Genomic Workflow Solutions
BioInformatics LLC has recognized Bio-Rad with two prestigious Life Science Industry Awards for “Best Protein Expression & Analysis Products” and “Best Workflow Solutions — Genomics.”
Monday, January 19, 2015
Droplet Digital PCR Enables Measurement of Potential Cancer Survival Biomarker
Study paves the way for further research into the role of TIL quantification in immunotherapy and as a cancer survival predictor.
Tuesday, December 10, 2013
A Brief RT-qPCR “Field Guide” for MIQE Adherence
Bio-Rad’s Sean Taylor and Eli Mrkusich have published a practical guide for MIQE compliance.
Monday, December 09, 2013
Bio-Rad Announces Mini-Protean Tetra Cell Users Website
The new online social networking site is created for current and new Bio-Rad Mini-PROTEAN® system users.
Wednesday, August 20, 2008
Bio-Rad Launches new Website for Researchers Transfecting Mammalian Cells
Bio-Rad adds ‘Gene Transfer Protocols’ to its Gene Expression Gateway Web-site, dedicated to researchers working with mammalian cells.
Friday, June 27, 2008
Testing Results of the BioOdyssey Calligrapher MiniArrayer for Clinical Research Available
Bio-Rad announces the availability of a technical bulletin describing results of the testing of the BioOdyssey™ Calligrapher™ MiniArrayer.
Thursday, November 08, 2007
Scientific News
Big Genetics in BC: The American Society for Human Genetics 2016 Meeting
Themes at this year's meeting ranged from the verification, validation, and sharing of data, to the translation of laboratory findings into actionable clinical results.
Cancer Genetics: Key to Diagnosis, Therapy
When applied judiciously, cancer genetics directs caregivers to the right drug at the right time, while sparing patients of unnecessary or harmful treatments.
Accelerating the Detection of Foodborne Bacterial Outbreaks
The speed of diagnosis of foodborne bacterial outbreaks could be improved by a new technique developed by researchers at the Georgia Institute of Technology.
Top 10 Life Science Innovations of 2016
2016 has seen the release of some truly innovative products. To help you digest these developments, The Scientist have listed their top picks for the year.
Scientists Identify Unique Genomic Features in Testicular Cancer
The findings may shed light on factors in other cancers that influence their sensitivity to chemotherapy.
Secret Phenotypes: Disease Devils in Invisible Details
Algorithmic deep phenotyping exposes masses of hidden traits and possible subtle genetic connections relevant to unseen influences on disease.
Cracking the Code of a Deadly Virus
Researchers have exploited weaknesses in VEEV's genetic code, creating a far less deadly variant.
Hunting the Missing Link Between Genetics and the Environment
The International Phenome Centre Network (IPCN) works to transform healthcare through phenomics - the dynamic interactions between our genes and our environment.
Repurposing Genes for Brain Development
Mammalian bone gene may be repurposed to promote cognition in humans.
Enhancing CRISPR to Explore Further
Researchers have developed sOPTiKO, a more efficient and controllable CRISPR genome editing platform.
Skyscraper Banner

SELECTBIO Market Reports
Go to LabTube
Go to eposters
 
Access to the latest scientific news
Exclusive articles
Upload and share your posters on ePosters
Latest presentations and webinars
View a library of 1,800+ scientific and medical posters
3,900+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
5,300+ scientific videos
Close
Premium CrownJOIN TECHNOLOGY NETWORKS PREMIUM FOR FREE!