Comparative Analysis and Optimization of Different DNA Extraction Protocols in Satureja khuzistanica

Doosty, Behrooz; Drikvand, Reza Mir; Salahvarzi, Elham; Amiri, Hamzeh; Hadian, Javad

doi:10.5539/ijb.v4n4p111

Cited by 3 publications

(3 citation statements)

References 17 publications

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“…The modified CTAB-SDS method included the use of additional steps of sample extraction in phenol-chloroform and chloroform-isoamyl alcohol. These repeated steps facilitated the removal of the secondary metabolites and essential oils (Doosty et al, 2012) hence, leading to the extraction of highquality DNA for genetic studies.…”

Section: Dna Extraction Methodsmentioning

confidence: 99%

“…The metabolites cause shearing of the DNA or bind to taq polymerase; hence, inhibiting the polymerase chain reaction process (Sudan et al, 2017). Standard techniques of DNA extraction in plants are ineffective in isolating DNA from species with high content of secondary metabolites (Doosty et al, 2012). Therefore, optimisation of DNA protocols for individual plant species is crucial for plants with high phytochemical contents.…”

Section: Introductionmentioning

confidence: 99%

“…The efficiency of DNA extraction protocols for plants with high contents of secondary metabolites varies from one species to another. Doosty et al (2012) examined the efficiency of four published protocols of DNA isolation from a medicinal plant Satureja khuzistanica. These included Murry and Thompson (1980), Dellaporta (1983), Doyle and Doyle (1990) and Kang and Yang (2004).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Optimised germination protocol for jackfruit seeds and evaluation of methods for extraction of DNA suitable for genetic analysis

Ojwang¹,

Adan²,

Nyaboga³

et al. 2022

Afr. Crop Sci. J.

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Jackfruit (Artocarpus heterophyllus) is an underutilised plant that is promising in curbing food and nutritional security in sub-Saharan Africa. However, high level of secondary metabolites in its tissues significantly hampers its genetic characterisation for breeding purposes. Primarily, the compounds react with DNA during the extraction process, thus reducing its yield and quality. The utilisation of leaves from jackfruit seedlings is a potentially effective approach of addressing the challenge, however, limited information is available on efficient jackfruit seed germination procedures. Elucidating effective methods of jackfruit seed germination, and optimising protocols for DNA extraction is crucial in promoting its genetic characterisation studies for identification of superior varieties for propagation. The objective of this study was to evaluate methods of jackfruit seed germination, and DNA extraction procedures using jackfruit leaves. Pre-treatment of seeds with 3% hydrogen peroxide was effective in enhancing seed germination within a short time, compared to distilled water and 3% hydrochloric acid. We optimised a DNA extraction technique by combining CTAB-SDS based approach with an enhanced solvent extraction method. The technique yielded high quantity and quality of DNA from jackfruit leaves, which was appropriate for downstream polymerase chain reaction analysis. The sequence-related amplified polymorphism (SRAP) and simple sequence repeat (SSR) amplifications confirmed the effectiveness of the optimised CTAB-SDS based protocol for extraction of high quality DNA.

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Section: Dna Extraction Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Optimised germination protocol for jackfruit seeds and evaluation of methods for extraction of DNA suitable for genetic analysis

Ojwang¹,

Adan²,

Nyaboga³

et al. 2022

Afr. Crop Sci. J.

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Effect of DNA extraction in the Rosa canina L. identification under different processing temperature

Žiarovská

Kyseľ

Cimermanová

et al. 2017

Potr. S. J. F. Sci.

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Rosa canina, L. is widely used for medicinal purposes as well as in food industry where it is a valuable source, bioactive compounds and natural colorants. Actually, no specific method is recommended as suitable one for DNA extraction from rose hips. The aim of the study was to compare three commercial and three non-commercial methods to extract total genomic DNA from rose hips hyphanthium. Four methods are based on the precipitation in principle and two methods are based on resin-binding. Extracted DNA was proved for the effectivity in following PCR. In total, six different DNA isolations was performed for differently heat processes rose hips -fresh hyphanthium, 2-weeks frozen hyphanthium, dried hyphanthium (50 °C) and boiled hyphanthium (100 °C). The amplification parameters of 500 bp chloroplast gene amplicon were evaluated. Obtained amounts of extracted DNA was very variable not only for every individual method used but for individual treatment of samples, too. In general, non-commercial method provided higher amount of extracted DNA, but the A260/280 ratio was lower. When regarding the processing treatment of the samples, high differences were found among the samples untreated by heat and those that were dried or boiled for three of the used extraction methods. All the samples were positive for amplification, but different amounts of amplified product were obtained. The comparison of data for concentrations of extracted DNA and concentrations of amplified product showed large differences when regarding the achieved purity of DNA in extraction.

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Comparative Analysis of Six DNA Extraction Methods in Cowpea (Vigna unguiculata L.Walp)

Tan¹,

Huang²,

Tie³

et al. 2013

JAS

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High quality DNA extractions are a prerequisite for genetic studies of a variety of plants including cowpea (Vigna unguiculata). Nowadays, there are a great number of plant DNA extraction methods, and commercially available extraction kits are also becoming more and more popular. It appears that different procedures work best for different plant groups. Thus in the genetic studies of cowpea, which DNA extraction method to choose becomes a concern. To solve this problem, five classic plant DNA isolation methods, including three CTAB methods and two SDS methods, were compared and evaluated while isolation using a commercial kit was also undertaken. The DNA extracted by these six methods from two-week-old cowpea seedlings were analyzed according to their cost and time, yield, purity, integrity, and functionality in restriction endonuclease digestion and PCR (polymerase chain reaction) based downstream analysis. After the evaluation, one most suitable method, described by Dellaporta et al. (1983) was selected and chosen for isolating DNA from young leaves of cowpea seedlings. The cost and time required in this method was relatively low. In addition, the quantity and the quality of the DNA extracted by this method were high enough to perform hundreds of PCR-based reactions.

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Comparative Analysis and Optimization of Different DNA Extraction Protocols in Satureja khuzistanica

Cited by 3 publications

References 17 publications

Optimised germination protocol for jackfruit seeds and evaluation of methods for extraction of DNA suitable for genetic analysis

Optimised germination protocol for jackfruit seeds and evaluation of methods for extraction of DNA suitable for genetic analysis

Effect of DNA extraction in the Rosa canina L. identification under different processing temperature

Comparative Analysis of Six DNA Extraction Methods in Cowpea (Vigna unguiculata L.Walp)

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