An improved method for efficient isolation and purification of genomic DNA from filamentous cyanobacteria belonging to genera Anabaena, Nodularia and Nostoc

Kaczyńska, Angelika; Łoś, Marcin; Węgrzyn, Grzegorz

doi:10.2478/s13545-013-0058-y

Cited by 7 publications

(24 citation statements)

References 12 publications

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“…Sonication, which has been trialled in previous works (Srivatava et al . ; Kaczyńska, et al ., ), showed promising results, but it is a time‐consuming step compared with bead beating and can also be a source of cross contamination between samples. Sonication baths are not as efficient as sonicating the sample directly with a probe, limiting the utility of sonication for DNA extraction.…”

Section: Discussionmentioning

confidence: 99%

DNA extraction from benthic Cyanobacteria: comparative assessment and optimization

et al. 2016

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These commercial kits allow greater recovery and the elimination of dangerous chemicals for DNA extraction, making them the method of choice for the isolation of DNA from benthic mats. They also facilitate the extraction of DNA from benthic Cyanobacteria, which can help to improve the characterization of Cyanobacteria in environmental studies using qPCRs or population composition analysis using next-generation sequencing.

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Section: Discussionmentioning

confidence: 99%

DNA extraction from benthic Cyanobacteria: comparative assessment and optimization

et al. 2016

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“…Cyanobacterial genomic DNA was isolated and purified as described previously [[13]], and 20 μg of DNA from each isolation experiment were used for library construction. The final product of each construction procedure was 1 ml of phage lysate which was then used to infect E. coli T1 R host strain (provided in CopyControl Fosmid Library Production Kit).…”

Section: Methodsmentioning

confidence: 99%

“…If yes, this strategy might be used in subsequent works for isolation and characterization of compounds revealing particular activities, which would be significantly facilitated by availability of specific clones, eliminating potential problems with the amount of tested compounds and reproducibility of results. Efficient construction of the libraries was possible, between others, due to employment of a recently optimized method for isolation and purification of genomic DNA from filamentous cyanobacteria, suitable for construction of genomic libraries [[ 13 ]]. Such procedures are challenging due to production by cyanobacteria of large amounts of cellulose, pectins, murein and xylose, which are components of the cell wall.…”

Section: Introductionmentioning

confidence: 99%

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The use of fosmid metagenomic libraries in preliminary screening for various biological activities

et al. 2014

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BackgroundIt is generally believed that there are many natural sources of as yet unknown bioactive compounds with a high biotechnological potential. However, the common method based on the use of cell extracts in the preliminary screening for particular molecules or activities is problematic as amounts of obtained compounds may be low, and such experiments are hardly reproducible. Therefore, the aim of this work was to test whether a novel strategy to search for previously unknown biological activities can be efficient. This strategy is based on construction of metagenomic libraries and employment of Escherichia coli strains as cell factories producing compounds of properties potentially useful in biotechnology.ResultsThree cyanobacterial metagenomic libraries were constructed in the fosmid system. The libraries were screened for various biological activities. Extracts from selected E. coli clones bearing constructs with fragments of cyanobacterial genomes revealed antimicrobial or anticancer activities. Interestingly, stimulation of growth of host bacteria bearing particular plasmids with certain cyanobacterial genes was detected, suggesting a potential possibility for improvement of E. coli cultivation during biotechnological production. The most interesting plasmids were sequenced, and putative mechanisms of biological effects caused by cyanobacterial gene products are discussed.ConclusionsThe strategy of exploring cyanobacteria as sources of bioactive compounds, based on E. coli cell factories producing compounds due to expression of genes from metagenomic libraries, appears to be effective.

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“…The DNA was purified by a chloroform wash and precipitated with isopropanol. Genomic DNA was eluted in 10 mM TRIS-HCl, pH 8 (Kumar et al, 2010;Kumar et al, 2013). The complete sequence of the 16S rRNA gene was amplified using primers 16S 8F (5'AGA GTT TGA TCM TGG CTC AG3') (Okibeet al, 2003) and 16S 1492R (5'GGT TAC CTT GTT ACG ACT T3') (Paramasivamet al, 2011).…”

Section: Molecular Analysismentioning

confidence: 99%

Actinobacteria Isolated From Mineral Ores in Peru

Hurtado¹,

Pacheco²,

Sheen³

et al. 2018

J microb biotech food sci

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A total of twenty-four isolates of actinobacteria from arsenopyrite, pyrite, polymetallic sulfides and magnetite from Peruvian mining zones have been used with the purpose of characterizing them by morphological, physiological and molecular studies for future biotechnological applications. 23 strains were identified as Streptomyces and1 as Actinomadura sp.95% and 70% of the strains were able to grow at pH 3.5 and pH 2, respectively. Only 19% of them were able to oxidize iron at pH 2. Also, they have been tested by growth on arsenopyrite tailings and Streptomyces sp. E1 and Streptomycesvariabilis AB5 leached 19.1 % and 15.5 % of arsenic, respectively, while the control without inoculum only showed 2.5% of leaching. Results indicate that these strains show important characteristics for leaching processes.

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An improved method for efficient isolation and purification of genomic DNA from filamentous cyanobacteria belonging to genera Anabaena, Nodularia and Nostoc

Cited by 7 publications

References 12 publications

DNA extraction from benthic Cyanobacteria: comparative assessment and optimization

DNA extraction from benthic Cyanobacteria: comparative assessment and optimization

The use of fosmid metagenomic libraries in preliminary screening for various biological activities

Actinobacteria Isolated From Mineral Ores in Peru

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