ArticleApplications in plant sciences2020
Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae.
Article in Applications in plant sciences, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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6 citing papers in PubMed, 13 citations in OpenAlex.
- Best practices for methylome characterization in novel species: a case study in the microalgae Microchloropsis.Communications biology · 2025Article
- Penisimplicins A and B: Novel Polyketide-Peptide Hybrid Alkaloids from the FungusMolecules (Basel, Switzerland) · 2024Article
- Simple and Effective Squash-PCR for Rapid Genotyping of Industrial Microalgae.Life (Basel, Switzerland) · 2024Article
- Genomic characterisation and ecological distribution ofFrontiers in microbiology · 2024Article
- Article
- Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae.Applications in plant sciences · 2020Article
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3 authors at 1 institution in 1 country.
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Abstract
premiseNew sequencing technologies have facilitated genomic studies in green microalgae; however, extracting high-quality DNA is often a bottleneck for long-read sequencing. METHODS AND
resultsHere, we present a low-cost, highly transferrable method for the extraction of high-molecular-weight (HMW), high-purity DNA from microalgae. We first determined the effect of sample preparation on DNA quality using three homogenization methods: manual grinding using a mini-pestle, automatic grinding using a vortex adapter, and grinding in liquid nitrogen. We demonstrated the versatility of grinding in liquid nitrogen followed by a modified cetyltrimethylammonium bromide (CTAB) extraction across a suite of aquatic- and desert-evolved algal taxa. Finally, we tested the protocol's robustness by doubling the input material to increase yield, producing per sample up to 20 μg of high-purity DNA longer than 21.2 kbp.
conclusionsAll homogenization methods produced DNA within acceptable parameters for purity, but only liquid nitrogen grinding resulted in HMW DNA. The optimization of cell lysis while minimizing DNA shearing is therefore crucial for the isolation of DNA for long-read genomic sequencing because template DNA length strongly affects read output and length.
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