Optimized DNA extraction and purification method from Alchemilla species using polyethylene glycol

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Shokatyari Samira; BEYGOM FAGHIR MARZIEH; KAZEMPOUR OSALOO SHAHROKH; Sohani Mohammad Mahdi

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Journal: Rostaniha Year:2020 | Volume:21 | Issue:2 (60) Page(s): 218-230

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Title

Optimized DNA extraction and purification method from Alchemilla species using polyethylene glycol

Author(s)

Shokatyari Samira | BEYGOM FAGHIR MARZIEH | KAZEMPOUR OSALOO SHAHROKH | Sohani Mohammad Mahdi | Issue Writer Certificate

Keywords

Nucleic acid purityQ1

phenolic compoundsQ1

polysaccharide compoundsQ1

secondary metabolitesQ1

spectrophotometryQ1

Abstract

Pure DNA is essential in various techniques of molecular biology and its extraction from plants to produce large amounts of secondary metabolites is a difficult task. Alchemilla is known to synthesize a large number of secondary metabolites which reduce the quality of the extracted DNA. This study, aimed to set up a method for high-quality DNA isolation from Alchemilla leaf. We examined three extraction methods. Furthermore, a comparison concerning price, simplicity, and security is carried out. We optimized a CTAB-based method using increasing the volume and concentration of CTAB buffer, lysis time, and cold incubation period, performing six times dilutions, and three times precipitations, adding polyethylene glycol, and removing toxic or expensive materials. The results showed that, 260/280 and 260/230 ratios of extracted DNA by the optimized method with the concentration of 595– 387 ng/µ L were 1. 75– 1. 82 and 1. 56– 1. 68, respectively. The quality of extracted DNA by this method was significantly higher (p < 0. 001 ) than that of other ways, so that all samples were positive for DNA, as assessed by electrophoresis and PCR. The optimized method was simple, effective, reproducible, relatively non-toxic, and inexpensive. The results revealed that, this method was successful in producing large amounts of high-quality amplifiable DNA.

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APA: Copy

Shokatyari, Samira, BEYGOM FAGHIR, MARZIEH, KAZEMPOUR OSALOO, SHAHROKH, & Sohani, Mohammad Mahdi. (2020). Optimized DNA extraction and purification method from Alchemilla species using polyethylene glycol. ROSTANIHA, 21(2 (60) ), 218-230. SID. https://sid.ir/paper/396737/en

Vancouver: Copy

Shokatyari Samira, BEYGOM FAGHIR MARZIEH, KAZEMPOUR OSALOO SHAHROKH, Sohani Mohammad Mahdi. Optimized DNA extraction and purification method from Alchemilla species using polyethylene glycol. ROSTANIHA[Internet]. 2020;21(2 (60) ):218-230. Available from: https://sid.ir/paper/396737/en

IEEE: Copy

Samira Shokatyari, MARZIEH BEYGOM FAGHIR, SHAHROKH KAZEMPOUR OSALOO, and Mohammad Mahdi Sohani, “Optimized DNA extraction and purification method from Alchemilla species using polyethylene glycol,” ROSTANIHA, vol. 21, no. 2 (60) , pp. 218–230, 2020, [Online]. Available: https://sid.ir/paper/396737/en

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