ArticleProceedings of the National Academy of Sciences of the United States of America2008
A comprehensive assay for targeted multiplex amplification of human DNA sequences.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
46 citing papers in PubMed.
- Virus Genome Sequences in the Blood of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome Patients.medRxiv : the preprint server for health sciences · 2025Article
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- Massively parallel DNA target capture using long adapter single stranded oligonucleotide (LASSO) probes assembled through a novel DNA recombinase mediated methodology.Biotechnology journal · 2022Article
- Sequencing Ultrarare Targets with Compound Nucleic Acid Cytometry.Analytical chemistry · 2021Article
- A multiplex platform for digital measurement of circular DNA reaction products.Nucleic acids research · 2020Article
- BART-Seq: cost-effective massively parallelized targeted sequencing for genomics, transcriptomics, and single-cell analysis.Genome biology · 2019Article
- A novel emulsion PCR method coupled with fluorescence spectrophotometry for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets.Scientific reports · 2019Article
- A CRISPR-Cas9-triggered strand displacement amplification method for ultrasensitive DNA detection.Nature communications · 2018Article
- Detection of 16S rRNA and KPC Genes from Complex Matrix Utilizing a Molecular Inversion Probe Assay for Next-Generation Sequencing.Scientific reports · 2018Article
- Cat-D: a targeted sequencing method for the simultaneous detection of small DNA mutations and large DNA deletions with flexible boundaries.Scientific reports · 2017Article
- ThermoAlign: a genome-aware primer design tool for tiled amplicon resequencing.Scientific reports · 2017Article
- Long-adapter single-strand oligonucleotide probes for the massively multiplexed cloning of kilobase genome regions.Nature biomedical engineering · 2017Article
- Sequence capture by hybridization to explore modern and ancient genomic diversity in model and nonmodel organisms.Nucleic acids research · 2016Review
- A Rapid, High-Quality, Cost-Effective, Comprehensive and Expandable Targeted Next-Generation Sequencing Assay for Inherited Heart Diseases.Circulation research · 2015Article
- Overview of high throughput sequencing technologies to elucidate molecular pathways in cardiovascular diseases.Circulation research · 2013Review
- Multiplex target capture with double-stranded DNA probes.Genome medicine · 2013Article
- Long-range genomic enrichment, sequencing, and assembly to determine unknown sequences flanking a known microRNA.PloS one · 2013Article
- Article
- Improved coverage and accuracy with strand-conserving sequence enrichment.Genome medicine · 2013Article
- Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders.Science (New York, N.Y.) · 2012Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
We developed a robust and reproducible methodology to amplify human sequences in parallel for use in downstream multiplexed sequence analyses. We call the methodology SMART (Spacer Multiplex Amplification Reaction), and it is based, in part, on padlock probe technology. As a proof of principle, we used SMART technology to simultaneously amplify 485 human exons ranging from 100 to 500 bp from human genomic DNA. In multiple repetitions, >90% of the targets were successfully amplified with a high degree of uniformity, with 70% of targets falling within a 10-fold range and all products falling within a 100-fold range of each other in abundance. We used long padlock probes (LPPs) >300 bases in length for the assay, and the increased length of these probes allowed for the capture of human sequences up to 500 bp in length, which is optimal for capturing most human exons. To engineer the LPPs, we developed a method that generates ssDNA molecules with precise ends, using an appropriately designed dsDNA template. The template has appropriate restriction sites engineered into it that can be digested to generate nucleotide overhangs that are suitable for lambda exonuclease digestion, producing a single-stranded probe from dsDNA. The SMART technology is flexible and can be easily adapted to multiplex tens of thousands of target sequences in a single reaction.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.