ArticleScientific reports2014
Long-range PCR in next-generation sequencing: comparison of six enzymes and evaluation on the MiSeq sequencer.
Article in Scientific reports, 2014. 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.
- Longrange PCR-based next-generation sequencing in pharmacokinetics and pharmacodynamics study of propofol among patients under general anaesthesia.Scientific reports · 2017Trial
- Capsid and genome damage are the leading inactivation mechanisms of aerosolized porcine respiratory coronavirus at different relative humidities.Applied and environmental microbiology · 2025Article
- Genetic analysis using long-read sequencing to overcome the difficulties inResearch and practice in thrombosis and haemostasis · 2025Article
- Efficient variant phasing utilizing a replication cycle reaction system.Genetics in medicine open · 2025Article
- Enzyme Catalysis for Sustainable Value Creation Using Renewable Biobased Resources.Molecules (Basel, Switzerland) · 2024Review
- Targeted phasing of 2-200 kilobase DNA fragments with a short-read sequencer and a single-tube linked-read library method.Scientific reports · 2024Article
- Identifying the best PCR enzyme for library amplification in NGS.Microbial genomics · 2024Article
- CRISPR-Cas9-guided amplification-free genomic diagnosis for familial hypercholesterolemia using nanopore sequencing.PloS one · 2024Article
- GREPore-seq: A Robust Workflow to Detect Changes After Gene Editing Through Long-range PCR and Nanopore Sequencing.Genomics, proteomics & bioinformatics · 2023Article
- Lentiviral Transduction-based CRISPR/Cas9 Editing ofCurrent genomics · 2023Article
- Beyond assembly: the increasing flexibility of single-molecule sequencing technology.Nature reviews. Genetics · 2023Review
- Fast and efficient template-mediated synthesis of genetic variants.Nature methods · 2023Article
- Long-range PCR in next generation sequencing: A low cost approach for large & complex genes.The Indian journal of medical research · 2023Article
- Long-range PCR amplification-based targeted enrichment & next generation sequencing: A cost-effective testing strategy for lysosomal storage disorders.The Indian journal of medical research · 2023Article
- Improving Yields in Multi-analyte Extractions by Utilizing Post-homogenized Tissue Debris.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2023Article
- First mitochondrial genome-wide association study with metabolomics.Human molecular genetics · 2022Article
- Identification and analysis of deletion breakpoints in four Mohr-Tranebjærg syndrome (MTS) patients.Scientific reports · 2022Article
- Target Enrichment Approaches for Next-Generation Sequencing Applications in Oncology.Diagnostics (Basel, Switzerland) · 2022Review
- Human whole mitochondrial genome sequencing and analysis: optimization of the experimental workflow.Croatian medical journal · 2022Article
- High-speed rail model reveals the gene tandem amplification mediated by short repeated sequence in eukaryote.Scientific reports · 2022Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Long-range PCR remains a flexible, fast, efficient and cost-effective choice for sequencing candidate genomic regions in a small number of samples, especially when combined with next-generation sequencing (NGS) platforms. Several long-range DNA polymerases are advertised as being able to amplify up to 15 kb or longer genomic DNA. However, their real-world performance characteristics and their suitability for NGS remain unclear. We evaluated six long-range DNA polymerases (Invitrogen SequalPrep, Invitrogen AccuPrime, TaKaRa PrimeSTAR GXL, TaKaRa LA Taq Hot Start, KAPA Long Range HotStart and QIAGEN LongRange PCR Polymerase) to amplify three amplicons, with sizes of 12.9 kb, 9.7 kb, and 5.8 kb, respectively. Subsequently, we used the PrimeSTAR enzyme to amplify entire BRCA1 (83.2 kb) and BRCA2 (84.2 kb) genes from nine subjects and sequenced them on an Illumina MiSeq sequencer. We found that the TaKaRa PrimeSTAR GXL DNA polymerase can amplify almost all amplicons with different sizes and Tm values under identical PCR conditions. Other enzymes require alteration of PCR conditions to obtain optimal performance. From the MiSeq run, we identified multiple intronic and exonic single-nucleotide variations (SNVs), including one mutation (c.5946delT in BRCA2) in a positive control. Our study provided useful results for sequencing research focused on large genomic regions.
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