2020
DOI: 10.21519/0234-2758-2020-36-6-98-106
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Extraction of DNA from a Wide Range of Objects by Treatment with Ammonium Salts

Abstract: Currently, simple, rapid, and efficient techniques for DNA isolation from a wide range of organisms are in demand in biotechnology and bioinformatics. A key (and often limiting) step is the cell wall disruption and subsequent DNA extraction from the disintegrated cells. We have developed a new approach to DNA isolation from organisms with robust cell walls. The protocol includes the following steps: treatment of cells or tissue samples with ammonium acetate followed by cell lysis in low-salt buffer with the ad… Show more

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Cited by 4 publications
(2 citation statements)
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“…For genomic DNA extraction, the strain was grown on nutrient broth (5 g/liter yeast extract, 5 g/liter tryptone, 2.5 g/liter NaCl [pH 7.2]) at 30°C for 2 days. DNA was extracted using the method described in reference 5 , and a DNA fragment library was prepared using the KAPA HyperPlus kit according to the manufacturer’s recommendations. Sequencing was performed on the Illumina MiSeq system at NRC Kurchatov Institute (Moscow, Russia) using the MiSeq reagent kit v2, and 3,363,592 paired-end 251-bp reads were generated.…”
Section: Announcementmentioning
confidence: 99%
“…For genomic DNA extraction, the strain was grown on nutrient broth (5 g/liter yeast extract, 5 g/liter tryptone, 2.5 g/liter NaCl [pH 7.2]) at 30°C for 2 days. DNA was extracted using the method described in reference 5 , and a DNA fragment library was prepared using the KAPA HyperPlus kit according to the manufacturer’s recommendations. Sequencing was performed on the Illumina MiSeq system at NRC Kurchatov Institute (Moscow, Russia) using the MiSeq reagent kit v2, and 3,363,592 paired-end 251-bp reads were generated.…”
Section: Announcementmentioning
confidence: 99%
“…Before DNA isolation, the strains were cultured in MRS medium for 36 h at 28°C to 30°C under aerobic conditions. The genomic DNA was extracted as described previously ( 8 ). Fragment genomic libraries were prepared with a Nextera XT reagent kit (Illumina, San Diego, CA, USA) according to the manufacturer’s recommendations.…”
Section: Announcementmentioning
confidence: 99%