Satellite Banner
Next Gen Sequencing
Scientific Community
 
Become a Member | Sign in
Home>News>This Article
  News
Return

Population Genetics Unveils Reflex™ Method Sequencing Tool

Published: Tuesday, April 23, 2013
Last Updated: Tuesday, April 23, 2013
Bookmark and Share
Method used for simultaneously sequencing long contiguous DNA regions in thousands of samples.

Researchers at Population Genetics Technologies Ltd (Cambridge, UK) have developed and validated an innovative technology - Reflex™ - for efficient targeted sequencing of long DNA regions in large numbers of genomic DNA samples.

Targeted sequencing is used to study specific parts of a genome that may be involved in disease or other relevant clinical traits. Unveiling the role of these genomic regions usually requires interrogating long contiguous DNA sequences, such as a gene or genes, and doing so in many hundreds or thousands of people. Samples from these individuals can be pooled to take advantage of the high capacity of current sequencing platforms, but current targeting approaches require processing of each sample separately to generate the multiplicity of small fragments required for next generation sequencing. Reflex™ starts with pools of large genomic regions from hundreds or thousands of samples, performing fragmentation on the pool yet retaining the initial sample identity, thus greatly increasing the efficiency and decreasing the cost of targeted sequencing of contiguous genomic regions in large sample numbers.

This Reflex™ technology uses an intramolecular reaction to derive the shorter, sequencer-ready, daughter products from a pooled population of barcoded long-range PCR products while preserving the cognate DNA barcodes (Nucleic Acids Research, 2013, doi:10.1093/nar/gkt228). This allows the large targeted region from many thousands of samples to be processed simultaneously in a pool, while allowing the derived sequences to be matched back to each individual. The size of the targeted region depends on the desired design of the long-range PCR, but typically will span 7-10 kilobases.
The Reflex workflow enables uniform sequence coverage of long contiguous sequence targets in large numbers of samples at low cost on desktop next-generation sequencers. The method requires small amounts of input genomic DNA and can be used to target members of multi-gene families with high specificity. The technique is platform-agnostic, having been used successfully on Roche 454, Ion Torrent and Illumina platforms.

Current next-generation sequencing (NGS) platforms require that adaptors are added to the ends of short target DNA fragments to be sequenced. Adding a multiplex identifier (MID), a short DNA barcode that identifies the sample, with the sequencing adaptor allows multiple DNA samples to be processed in a single sequencing run. Typically, individual samples are prepared and then pooled at the sequencing step, requiring expensive and labor-intensive preparation methods: thus, for targeted re-sequencing, sample preparation costs dominate the overall cost.

Population Genetics CEO, Alan Schafer said that this issue motivated Population Genetics founder, Nobel Laureate Sydney Brenner, to invent a technique that can perform sample preparation on a pooled population of long ‘parent’ DNA fragments which are already appended with adaptors and MIDs, to generate smaller, sequencer-ready, ‘daughter’ amplicons that preserve the adaptors and MIDs.

“Many laboratories are interrogating the same genomic regions in many hundreds, if not thousands, of samples and can benefit from the sample-scale efficiencies of Reflex”. “When coupled with sequencers that allow an extra indexing run, the method can be used to simultaneously sequence thousands of samples in a single run”, he said. The company has already used the Reflex workflow in this way to extract and sequence a gene target from 3000 human genomic DNA samples as part of an on-going disease susceptibility collaboration.

Reflex technology also has the potential to generate long reads within and beyond each starting long range PCR product by propagating molecular identifiers across a contiguous region (in development at Population Genetics). The resulting data can inform haplotyping, genome phasing and RNA isoform identification using short-read NGS platforms, extending its value in providing coverage of clinically important genes and genomes.


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

Population Genetics Launches Populus Award 2012
Award offers financial and expert practical support for the completion of a multi-sample gene sequencing project using Population Genetics’ proprietary technologies.
Tuesday, October 16, 2012
Population Genetics Launches Funding Award for Novel Population Sequencing Studies
The Populus Award 2012 offers financial and expert practical support for the completion of a multi-sample gene sequencing project.
Monday, October 15, 2012
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.
Antibiotics Discovered While Sifting Through Human Microbiome
Researchers identify genes in a microbe’s genome that produce antibiotic compounds, then synthesize them without the need for bacterial culture.
A Genome-wide View of Human DNA Viruses
In this study, Duplex sequencing was used to accurately analyse the genome-wide rate of spontaneous mutation of human adenovirus C5 (HAdv5).
Peer Review is in Crisis, But Should be Fixed, Not Abolished
After the time to get the science done, peer review has become the slowest step in the process of sharing studies, and some scientists have had enough.
Rare Immunodeficiency Yields Unexpected Insights
Scientists uncover previously unknown gene mutation revealing the role of a key molecule involved in immune cell development.
Illumina Contributes to ClinVar Database
The contribution includes variants of all classifications, from pathogenic to benign, identified during interpretation of whole genome sequences generated in the CLIA-certified, CAP-accredited Illumina Clinical Services Laboratory.
Agilent Presents Early Career Professor Award to Dr. Roeland Verhaak
JAX professor recognized for the development and implementation of workflows for the analysis of big-data from transcriptomics to next generation sequencing approaches.
Integrated Omics Analysis
Studying multi-omics promises to give a more holistic picture of the organism and its place in its ecosystem, however despite the complexities involved those within the field are optimistic.
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!