Genomic DNA Extraction Protocol for Artemisia Annua L. Without Using Liquid Nitrogen and Phenol

Agrawal, Ankit; Sharma, Anamika; Shukla, Narmada Prasad

doi:10.3126/ijasbt.v4i4.15788

Cited by 4 publications

(2 citation statements)

References 6 publications

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“…High polyphenol, polysaccharide and other secondary metabolite content can interfere with DNA extraction and purification [16,17]. Especially difficult are the oxidized form of polyphenols, as they can covalently bind to DNA interfering with the purification steps in DNA extraction [18,19]. Also, polysaccharides can affect downstream digestion and amplification processes by inhibiting restriction enzyme and Taq polymerase activity, respectively.…”

Section: Introductionmentioning

confidence: 99%

An Improved Method for Efficient DNA Extraction from Grapevine

Öncü‐Öner

TEMEL

PAMAY

et al. 2023

International Journal of Life Sciences and Biotechnology

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Grapevine (Vitis vinifera L.) is one of the oldest and most important perennial crops worldwide which has been the subject of extensive genetic studies including gene mapping, genetic transformation, and DNA fingerprinting. Grapevines are rich in polysaccharides, polyphenolic compounds, and various secondary metabolites, many of which have significant importance in food, agrochemical, and pharmaceutical industries. While metabolites are one of the indicators of quality of grapevines, the presence of them makes grapevine one of the most difficult plants to extract DNA from. These metabolites not only affect DNA extraction procedures but also downstream reactions such as restriction digestion and PCR. Development of new genotyping techniques based on sequencing such as genotyping by sequencing (GBS) requires high-quality DNA for digestion and sequencing. To date, several protocols have been developed for DNA extraction from grapevine. In this study, three different protocols with modifications were compared for DNA extraction performance from grapevine leaves from four different cultivars. Efficiencies of these methods were determined by extracted DNA’s quantity and quality. To confirm the suitability for GBS, extracted DNA was digested with restriction enzymes. Although all protocols were based on the traditional CTAB method, they resulted in different DNA yield and restriction digestion efficiency. The modified protocol including PVP-40 and ß-mercaptoethanol was found to be the most efficient method to obtain high quality and quantity grapevine DNA that is amenable to restriction digestion.

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Section: Introductionmentioning

confidence: 99%

An Improved Method for Efficient DNA Extraction from Grapevine

Öncü‐Öner

TEMEL

PAMAY

et al. 2023

International Journal of Life Sciences and Biotechnology

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“…During cell lysis, nucleic acids come into contact with these polysaccharides in the oxidized form. The polyphenols bind covalently and irreversibly to proteins and nucleic acids resulting in a brown gelatinous material (Agrawal et al, 2016). This reduces the yield and purity of DNA.…”

Section: Introductionmentioning

confidence: 99%

Comparison and optimization for DNA extraction of okra (Abelmoschus esculentus L. Moench)

Kidane¹,

Kalousová²,

Demissie³

2020

Afr. J. Biotechnol.

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The extraction of high-quality DNA from okra (Abelmoschus esculentus L. Moench) is notoriously troublesome due to the high contents of polysaccharides, polyphenols, and different secondary metabolites. We have tested seven extraction buffers on silica dried okra leaves. Here, we describe a simple, rapid and modified procedure for high-quality DNA extraction from okra, which is amenable for downstream analyses. In contrast to Cetyl-trimethyl-ammonium bromide (CTAB) methods, the described procedure is rapid, omits the use of liquid nitrogen, phenol, PVP-10, and chloroform. It also uses inexpensive and less hazardous reagents and requires only ordinary laboratory equipment. The procedure employed a high concentration of Sodium dodecyl sulphate (SDS) to rid the problems associated with polysaccharides and polyphenols. The average yield was between 36 and 45 μg of total DNA from 90 mg of dried leaf weight. The DNA is adequate for molecular analysis of okra, such as genetic mapping or marker-assisted plant breeding. This protocol can be performed in as little as 3 h and may be adapted to high-throughput DNA isolation.

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