ArticlePloS one2012
An improved chloroplast DNA extraction procedure for whole plastid genome sequencing.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 96 papers.
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96 citing papers in PubMed.
- The Complete Chloroplast Genome ofGenes · 2026Article
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- Comprehensive plastid phylogenomics reveal the phylogeny ofPhytoKeys · 2026Article
- The Complete Chloroplast Genomes of ThreeCurrent issues in molecular biology · 2025Article
- Carboxylation and Oxygenation Kinetics and Large Subunit (rbcL) DNA Sequences for Rubisco From Two Ecotypes of Plantago lanceolata L. That Are Native to Sites Differing in Atmospheric COPlant, cell & environment · 2025Article
- The complete chloroplast genome sequences of three Cypripedium species and their phylogenetic analysis.Scientific reports · 2025Article
- Comparative chloroplast genomics provides insights into the phylogenetic relationships and evolutionary history for Actinidia species.Scientific reports · 2025Article
- Characterization of the complete chloroplast genome and comparative analysis of the phylogeny and codon usage bias of three Yunnan wild rice species.Frontiers in plant science · 2025Article
- Extraction and analysis of high-quality chloroplast DNA with reduced nuclear DNA for medicinal plants.BMC biotechnology · 2024Article
- A novel in-situ-process technique constructs whole circular cpDNA library.Plant methods · 2024Article
- Kazakhstan tulips: comparative analysis of complete chloroplast genomes of four local and endangered species of the genusFrontiers in plant science · 2024Article
- The complete chloroplast genome of Cicer reticulatum and comparative analysis against relative Cicer species.Scientific reports · 2023Article
- Complete chloroplast genome of Lens lamottei reveals intraspecies variation among with Lens culinaris.Scientific reports · 2023Article
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- Comparative plastome genomics, taxonomic delimitation and evolutionary divergences of Tetraena hamiensis var. qatarensis and Tetraena simplex (Zygophyllaceae).Scientific reports · 2023Article
- Comparison of five DNA extraction methods in three medicinal plants:Molecular biology research communications · 2023Article
- Article
- Complete chloroplast genome sequencing and comparative analysis of threatened dragon trees Dracaena serrulata and Dracaena cinnabari.Scientific reports · 2022Article
- Complete chloroplast genome sequence of Lens ervoides and comparison to Lens culinaris.Scientific reports · 2022Article
- Article
36 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundChloroplast genomes supply valuable genetic information for evolutionary and functional studies in plants. The past five years have witnessed a dramatic increase in the number of completely sequenced chloroplast genomes with the application of second-generation sequencing technology in plastid genome sequencing projects. However, cost-effective high-throughput chloroplast DNA (cpDNA) extraction becomes a major bottleneck restricting the application, as conventional methods are difficult to make a balance between the quality and yield of cpDNAs. METHODOLOGY/PRINCIPAL
findingsWe first tested two traditional methods to isolate cpDNA from the three species, Oryza brachyantha, Leersia japonica and Prinsepia utihis. Both of them failed to obtain properly defined cpDNA bands. However, we developed a simple but efficient method based on sucrose gradients and found that the modified protocol worked efficiently to isolate the cpDNA from the same three plant species. We sequenced the isolated DNA samples with Illumina (Solexa) sequencing technology to test cpDNA purity according to aligning sequence reads to the reference chloroplast genomes, showing that the reference genome was properly covered. We show that 40-50% cpDNA purity is achieved with our method.
conclusionHere we provide an improved method used to isolate cpDNA from angiosperms. The Illumina sequencing results suggest that the isolated cpDNA has reached enough yield and sufficient purity to perform subsequent genome assembly. The cpDNA isolation protocol thus will be widely applicable to the plant chloroplast genome sequencing projects.
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