ArticlePloS one2016
Improved PCR Amplification of Broad Spectrum GC DNA Templates.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 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
11 citing papers in PubMed.
- Optimization of PCR amplification and sequencing of GC-rich protein-coding region of CEBPA (CCAAT/enhancer binding protein alpha) in acute myeloid leukemia patients using betaine.Molecular biology reports · 2026Article
- Cell-Free DNA-Based Theranostics for Inflammatory Disorders.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- The Appalbees menu: a multiyear, multilocus metagenetic assessment of pollen foraging by AppalachianPeerJ · 2026Article
- DeepCodon: A deep learning codon-optimization model to enhance protein expression.Biodesign research · 2025Article
- Development of a loop-mediated isothermal amplification method for the rapid detection of Clostridium botulinum serotypes E and F.Molecular biology reports · 2024Article
- CleanUpRNAseq: An R/Bioconductor Package for Detecting and Correcting DNA Contamination in RNA-Seq Data.Biotech (Basel (Switzerland)) · 2024Article
- Simultaneous inhibition of DNA-PK and Polϴ improves integration efficiency and precision of genome editing.Nature communications · 2023Article
- Development of a program for in silico optimized selection of oligonucleotide-based molecular barcodes.PloS one · 2021Article
- Consensus PCR protocols for the detection of amphibian herpesviruses (Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2020Article
- Article
- Development and application of fluorescent loop mediated isothermal amplification technique to detect3 Biotech · 2019Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Many applications in molecular biology can benefit from improved PCR amplification of DNA segments containing a wide range of GC content. Conventional PCR amplification of DNA sequences with regions of GC less than 30%, or higher than 70%, is complex due to secondary structures that block the DNA polymerase as well as mispriming and mis-annealing of the DNA. This complexity will often generate incomplete or nonspecific products that hamper downstream applications. In this study, we address multiplexed PCR amplification of DNA segments containing a wide range of GC content. In order to mitigate amplification complications due to high or low GC regions, we tested a combination of different PCR cycling conditions and chemical additives. To assess the fate of specific oligonucleotide (oligo) species with varying GC content in a multiplexed PCR, we developed a novel method of sequence analysis. Here we show that subcycling during the amplification process significantly improved amplification of short template pools (~200 bp), particularly when the template contained a low percent of GC. Furthermore, the combination of subcycling and 7-deaza-dGTP achieved efficient amplification of short templates ranging from 10-90% GC composition. Moreover, we found that 7-deaza-dGTP improved the amplification of longer products (~1000 bp). These methods provide an updated approach for PCR amplification of DNA segments containing a broad range of GC content.
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Registered trials
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