ArticleNucleic acids research2005
Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments.
Article in Nucleic acids research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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Who cites it
57 citing papers in PubMed.
- A ligase-based toolbox for research and diagnostics in molecular medicine.Nucleic acids research · 2026Review
- Ultra-sensitive monitoring of leukemia patients using superRCA mutation detection assays.Nature communications · 2022Article
- Dealing with Pseudogenes in Molecular Diagnostics in the Next Generation Sequencing Era.Methods in molecular biology (Clifton, N.J.) · 2021Review
- Article
- A novel emulsion PCR method coupled with fluorescence spectrophotometry for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets.Scientific reports · 2019Article
- Limited reverse transcriptase activity of phi29 DNA polymerase.Nucleic acids research · 2018Article
- Rapid and highly-specific generation of targeted DNA sequencing libraries enabled by linking capture probes with universal primers.PloS one · 2018Article
- Sensitive and inexpensive digital DNA analysis by microfluidic enrichment of rolling circle amplified single-molecules.Nucleic acids research · 2017Article
- Targeted DNA sequencing and in situ mutation analysis using mobile phone microscopy.Nature communications · 2017Article
- From Genomes to GENE-omes: Exome Sequencing Concept and Applications in Crop Improvement.Frontiers in plant science · 2017Review
- Clonal rolling circle amplification for on-chip DNA cluster generation.Biology methods & protocols · 2017Article
- PrimerPooler: automated primer pooling to prepare library for targeted sequencing.Biology methods & protocols · 2017Article
- Microbial Ecology: Where are we now?Postdoc journal : a journal of postdoctoral research and postdoctoral affairs · 2016Article
- Clinical Versus Research Sequencing.Cold Spring Harbor perspectives in medicine · 2016Review
- Single-locus enrichment without amplification for sequencing and direct detection of epigenetic modifications.Molecular genetics and genomics : MGG · 2016Article
- Clinical exome sequencing in neurogenetic and neuropsychiatric disorders.Annals of the New York Academy of Sciences · 2016Review
- Phenotypic heterogeneity in monogenic diabetes: the clinical and diagnostic utility of a gene panel-based next-generation sequencing approach.Molecular genetics and metabolism · 2014Article
- Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.Nature protocols · 2014Article
- Article
- Methods for the preparation of large quantities of complex single-stranded oligonucleotide libraries.PloS one · 2014Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We present a method to specifically select large sets of DNA sequences for parallel amplification by PCR using target-specific oligonucleotide constructs, so-called selectors. The selectors are oligonucleotide duplexes with single-stranded target-complementary end-sequences that are linked by a general sequence motif. In the selection process, a pool of selectors is combined with denatured restriction digested DNA. Each selector hybridizes to its respective target, forming individual circular complexes that are covalently closed by enzymatic ligation. Non-circularized fragments are removed by exonucleolysis, enriching for the selected fragments. The general sequence that is introduced into the circularized fragments allows them to be amplified in parallel using a universal primer pair. The procedure avoids amplification artifacts associated with conventional multiplex PCR where two primers are used for each target, thereby reducing the number of amplification reactions needed for investigating large sets of DNA sequences. We demonstrate the specificity, reproducibility and flexibility of this process by performing a 96-plex amplification of an arbitrary set of specific DNA sequences, followed by hybridization to a cDNA microarray. Eighty-nine percent of the selectors generated PCR products that hybridized to the expected positions on the array, while little or no amplification artifacts were observed.
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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.