ArticleNucleic acids research2011
Ice-COLD-PCR enables rapid amplification and robust enrichment for low-abundance unknown DNA mutations.
Article in Nucleic acids research, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 107 citations in OpenAlex.
- Combining E-ice-COLD-PCR and Pyrosequencing with Di-Base Addition (PDBA) Enables Sensitive Detection of Low-Abundance Mutations.Applied biochemistry and biotechnology · 2024Article
- Target Enrichment Approaches for Next-Generation Sequencing Applications in Oncology.Diagnostics (Basel, Switzerland) · 2022Review
- Mutation enrichment in human DNA samples via UV-mediated cross-linking.Nucleic acids research · 2022Article
- Limitations and opportunities of technologies for the analysis of cell-free DNA in cancer diagnostics.Nature biomedical engineering · 2022Review
- Selective multiplexed enrichment for the detection and quantitation of low-fraction DNA variants via low-depth sequencing.Nature biomedical engineering · 2021Article
- Establishment of multiplex allele-specific blocker PCR for enrichment and detection of 4 commonAnnals of translational medicine · 2020Article
- A nuclease-polymerase chain reaction enables amplification of probes used for capture-based DNA target enrichment.Nucleic acids research · 2019Article
- Denaturation-Enhanced Droplet Digital PCR for Liquid Biopsies.Clinical chemistry · 2018Article
- A Universal Analysis Pipeline of Hybrid Capture-Based Targeted Sequencing Data with Unique Molecular Indexes (UMIs).Genomics & informatics · 2018Article
- Evaluation of droplet digital PCR and next generation sequencing for characterizing DNA reference material for KRAS mutation detection.Scientific reports · 2018Article
- Efficient identification of SNPs in pooled DNA samples using a dual mononucleotide addition-based sequencing method.Molecular genetics and genomics : MGG · 2017Article
- Multiplexed Elimination of Wild-Type DNA and High-Resolution Melting Prior to Targeted Resequencing of Liquid Biopsies.Clinical chemistry · 2017Article
- Materials and microfluidics: enabling the efficient isolation and analysis of circulating tumour cells.Chemical Society reviews · 2017Review
- COLD-PCR Technologies in the Area of Personalized Medicine: Methodology and Applications.Molecular diagnosis & therapy · 2017Review
- Methylation-sensitive enrichment of minor DNA alleles using a double-strand DNA-specific nuclease.Nucleic acids research · 2017Article
- Multiplexed enrichment of rare DNA variants via sequence-selective and temperature-robust amplification.Nature biomedical engineering · 2017Article
- DMSO Increases Mutation Scanning Detection Sensitivity of High-Resolution Melting in Clinical Samples.Clinical chemistry · 2015Article
- Enhanced ratio of signals enables digital mutation scanning for rare allele detection.The Journal of molecular diagnostics : JMD · 2015Article
- Single-tube, highly parallel mutation enrichment in cancer gene panels by use of temperature-tolerant COLD-PCR.Clinical chemistry · 2015Article
- Extendable blocking probe in reverse transcription for analysis of RNA variants with superior selectivity.Nucleic acids research · 2015Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Identifying low-abundance mutations within wild-type DNA is important in several fields of medicine, including cancer, prenatal diagnosis and infectious diseases. However, utilizing the clinical and diagnostic potential of rare mutations is limited by sensitivity of the molecular techniques employed, especially when the type and position of mutations are unknown. We have developed a novel platform that incorporates a synthetic reference sequence within a polymerase chain reaction (PCR) reaction, designed to enhance amplification of unknown mutant sequences during COLD-PCR (CO-amplification at Lower Denaturation temperature). This new platform enables an Improved and Complete Enrichment (ice-COLD-PCR) for all mutation types and eliminates shortcomings of previous formats of COLD-PCR. We evaluated ice-COLD-PCR enrichment in regions of TP53 in serially diluted mutant and wild-type DNA mixtures. Conventional-PCR, COLD-PCR and ice-COLD-PCR amplicons were run in parallel and sequenced to determine final mutation abundance for a range of mutations representing all possible single base changes. Amplification by ice-COLD-PCR enriched all mutation types and allowed identification of mutation abundances down to 1%, and 0.1% by Sanger sequencing or pyrosequencing, respectively, surpassing the capabilities of other forms of PCR. Ice-COLD-PCR will help elucidate the clinical significance of low-abundance mutations and our understanding of cancer origin, evolution, recurrence-risk and treatment diagnostics.
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Registered trials
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