ArticleFrontiers in microbiology2023
Synthetic oligonucleotides as quantitative PCR standards for quantifying microbial genes.
Article in Frontiers in microbiology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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12 citing papers in PubMed, 20 citations in OpenAlex.
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- Development and validation of a quantitative pcr array assay for the detection of viral sequences in breast cancer.Journal, genetic engineering & biotechnology · 2026Article
- Optimization of Serum-Free Culture Medium for Theileria annulata-Transformed Cell Lines: A Comparative Analysis.Acta parasitologica · 2026Article
- Elevated graminoid cover co-occurs with Ascomycota-dominated soils in Longyearbyen, Svalbard.FEMS microbiology ecology · 2026Article
- Biochemical engineering of 5hmdC-DNA using a Tet3 double-mutant.Computational and structural biotechnology journal · 2026Article
- Article
- Identification of microbial groups and functional genes associated with greenhouse gas emissions in prawn farms.Scientific reports · 2025Article
- Development of functional markers and expression analysis for a Peroxidase gene TaPod-A3 on chromosome 7AL in wheat.The plant genome · 2025Article
- Revealing the composition of bacterial communities in various oil-contaminated soils and investigating their intrinsic traits in hydrocarbon degradation.Scientific reports · 2025Article
- Environmental Heterogeneity Drives Ecological Differentiation in Vibrio Populations Across Subtropical Marine Habitats.Environmental microbiology · 2025Article
- Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Real-time quantitative PCR (qPCR) has been widely used to quantify gene copy numbers in microbial ecology. Despite its simplicity and straightforwardness, establishing qPCR assays is often impeded by the tedious process of producing qPCR standards by cloning the target DNA into plasmids. Here, we designed double-stranded synthetic DNA fragments from consensus sequences as qPCR standards by aligning microbial gene sequences (10-20 sequences per gene). Efficiency of standards from synthetic DNA was compared with plasmid standards by qPCR assays for different phylogenetic marker and functional genes involved in carbon (C) and nitrogen (N) cycling, tested with DNA extracted from a broad range of soils. Results showed that qPCR standard curves using synthetic DNA performed equally well to those from plasmids for all the genes tested. Furthermore, gene copy numbers from DNA extracted from soils obtained by using synthetic standards or plasmid standards were comparable. Our approach therefore demonstrates that a synthetic DNA fragment as qPCR standard provides comparable sensitivity and reliability to a traditional plasmid standard, while being more time- and cost-efficient.
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