scholarly journals Evaluation of economical and rapid method of plant DNA extraction for PCR analysis of different crops

2017 ◽  
Vol 9 (2) ◽  
pp. 866-870
Author(s):  
Sweta Sinha ◽  
Amarendra Kumar
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Oryza Sativa L ◽  
Pcr Amplification ◽  
Leaf Tissue ◽  
Pcr Analysis ◽  
Time Saving ◽  
Dna Extraction Method ◽  
Molecular Studies ◽  
Ctab Method

In the recent genomic era, polymerase chain reaction (PCR) has become a basic tool in molecular studies and the success of PCR depends upon the template DNA. PCR technique is quite robust and often unnecessary to extract high quality of DNA and hence crude DNA can be used as template for amplification. Therefore, we have evaluated NaOH-Tris DNA extraction method for PCR analysis because this is very simple, time saving and safe without the need to use expensive or rare materials and laboratory apparatus. This method only requires a small amount of leaf tissue, NaOH, Tris, micro tube and plastic pestle. The amplified PCR products showed clear, sharp and uniform bands gave similar results as compared with the modified CTAB method. The DNA obtained is crude contains impurities like protein, RNA but these impurities did not affect PCR amplification. This DNA extraction method is evaluated for brinjal (Solanummelongena L.), chilli (Capsicum annuum L.), rice (Oryza sativa L.) and tomato (Solanumlycopersicum L.) crop. Many other crop plants could also be amplified using the same DNA extraction method for molecular analysis of large samples. Thus, the use of NaOH-Tris method will allow researchers to obtain DNA from plant quickly for use in molecular studies.

scholarly journals A simplified DNA extraction method for PCR analysis of Camarotella spp.

2010 ◽  
Vol 53 (2) ◽  
pp. 249-252 ◽  
Author(s):  
Nadja Santos Vitória ◽  
José Luiz Bezerra ◽  
Karina Peres Gramacho
Keyword(s):  
Dna Extraction ◽  
Plant Pathogen ◽  
Extraction Method ◽  
Pcr Analysis ◽  
Coconut Tree ◽  
Dna Extraction Method ◽  
Simple Protocol ◽  
Molecular Studies

This work aimed to optimize an efficient and simple protocol for DNA extraction of Camarotella species, an obligate plant pathogen that cause verrucosis or "lixa" on coconut tree and other palms, facilitating the molecular studies of these biotrophic microorganisms. The method proposed enabled a fast, reproducible and reliable DNA extraction from Camarotella species.

scholarly journals A modified SDS-based DNA extraction method from raw soybean

2019 ◽  
Vol 39 (2) ◽  
Author(s):  
Yimiao Xia ◽  
Fusheng Chen ◽  
Yan Du ◽  
Chen Liu ◽  
Guanhao Bu ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Monitoring Methods ◽  
Detection Techniques ◽  
Dna Yield ◽  
Seed Flour ◽  
Dna Extraction Method ◽  
Lysis Buffer

Abstract Soybean is the most important genetically modified (GM) oilseed worldwide. Regulations relating to the approval of biotech soybean varieties and product labeling demand accurate and reliable detection techniques to screen for GM soya. High-quality extracted DNA is essential for DNA-based monitoring methods. Thus, four widely used protocols (SDS, CTAB, DP305, and DNeasy Plant Mini Kit) were compared in the present study to explore the most efficient DNA extraction method for raw soya matrix. The SDS-based method showed the highest applicability. Then crucial factors influencing DNA yield and purity, such as SDS lysis buffer component concentrations and organic compounds used to isolate DNA, were further investigated to improve the DNA obtained from raw soybean seeds, which accounts for the innovation of this work. As a result, lysis buffer (2% SDS (w/v), 150 mM NaCl, 50 mM Tris/HCl, 50 mM EDTA, pH 8.0) and organic reagents including chloroform/isoamyl alcohol (24:1, v/v) (C: I), isopropanol, and ethanol corresponding to the extraction and first and second precipitation procedures, respectively, were used in the optimized SDS method. The optimized method was verified by extracting approximately 2020–2444 ng DNA/mg soybean with A260/280 ratios of 1.862–1.954 from five biotech and non-biotech soybean varieties. Only 0.5 mg of soya was required to obtain enough DNA for PCR amplification using the optimized SDS-based method. These results indicate that the screening protocol in the present study achieves the highest suitability and efficiency for DNA isolation from raw soya seed flour.

scholarly journals Comparison of genomic DNA extraction methods to obtain high DNA quality from barnyard grass (Echinochloa colona)

2020 ◽  
Author(s):  
Romesh Kumar Salgotra ◽  
Bhagirath Singh Chauhan
Keyword(s):  
Essential Oils ◽  
Dna Extraction ◽  
Leaf Tissue ◽  
Extraction Methods ◽  
High Quality ◽  
Time Saving ◽  
Plant Leaf ◽  
Plant Parts ◽  
Molecular Studies ◽  
Leaf Tissues

Abstract Background: The study of weed genomics is important for the effective management of weeds to enhance crop yield. A rapid, inexpensive and high quality DNA extraction is needed for genomic and other molecular studies. Here, we describe the protocols for DNA extraction from two different parts of the Echinochloa colona plant using modified cetyltrimethylammonium bromide (CTAB) and a commercial kit.Results: In the study, it was observed that the DNA extracted from plant leaf tissues and dry seeds with a modified CTAB protocol was of good quality, with no contaminations of polysaccharides and essential oils. Quality of DNA extracted from dry seeds was comparable with that of plant leaves under both protocols. The extracted DNA from both plant parts was successfully amplified by PCR using the EPSPS microsatellite marker. Compared to the protocol of DNA extracted from leaf tissue, dry seeds will save time and other valuable resources. Moreover, the same protocols can be implemented for the extraction of high-quality DNA for molecular studies in other plant species where a large amount of polysaccharides, secondary metabolites and essential oils are present.Conclusions: Modified methods of DNA extraction from dry seeds are efficient and time-saving which can be used in genotypic and other molecular approaches. High-quality DNA can be isolated from plant leaf tissues using modified CTAB and commercial kits, however, DNA extracted from dry seeds will save time and other valuable resources.

A Simple DNA Extraction Method for PCR Amplification from Dry Seeds of Brassica napus

2007 ◽  
Vol 10 (7) ◽  
pp. 1122-1125 ◽  
Author(s):  
Li Maoteng ◽  
Liu Jianmin ◽  
Zhangyi . ◽  
Wang Pei ◽  
Gan Lu ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Dna Extraction ◽  
Extraction Method ◽  
Pcr Amplification ◽  
Dna Extraction Method

Development of an Efficient Fungal DNA Extraction Method To Be Used in Random Amplified Polymorphic DNA–PCR Analysis To Differentiate Cyclopiazonic Acid Mold Producers

2008 ◽  
Vol 71 (12) ◽  
pp. 2497-2503 ◽  
Author(s):  
BEATRIZ SÁNCHEZ ◽  
MAR RODRÍGUEZ ◽  
EVA M. CASADO ◽  
ALBERTO MARTÍN ◽  
JUAN J. CÓRDOBA
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Cyclopiazonic Acid ◽  
Proteinase K ◽  
Vacuum System ◽  
Pcr Analysis ◽  
Dna Extraction Method ◽  
Rapd Pcr

A variety of previously established mechanical and chemical treatments to achieve fungal cell lysis combined with a semiautomatic system operated by a vacuum pump were tested to obtain DNA extract to be directly used in randomly amplified polymorphic DNA (RAPD)–PCR to differentiate cyclopiazonic acid–producing and –nonproducing mold strains. A DNA extraction method that includes digestion with proteinase K and lyticase prior to using a mortar and pestle grinding and a semiautomatic vacuum system yielded DNA of high quality in all the fungal strains and species tested, at concentrations ranging from 17 to 89 ng/μl in 150 μl of the final DNA extract. Two microliters of DNA extracted with this method was directly used for RAPD-PCR using primer (GACA)4. Reproducible RAPD fingerprints showing high differences between producer and nonproducer strains were observed. These differences in the RAPD patterns did not differentiate all the strains tested in clusters by cyclopiazonic acid production but may be very useful to distinguish cyclopiazonic acid producer strains from nonproducer strains by a simple RAPD analysis. Thus, the DNA extracts obtained could be used directly without previous purification and quantification for RAPD analysis to differentiate cyclopiazonic acid producer from nonproducer mold strains. This combined analysis could be adaptable to other toxigenic fungal species to enable differentiation of toxigenic and nontoxigenic molds, a procedure of great interest in food safety.

scholarly journals A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis

2020 ◽  
Author(s):  
Wei Hu ◽  
J. Clark Lagarias
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Method ◽  
Low Cost ◽  
Plant Molecular Biology ◽  
Pcr Analysis ◽  
High Quality ◽  
Plant Dna ◽  
Genomic Dna Isolation ◽  
Dna Extraction Method

AbstractBackgroundConsistent isolation of high quality plant genomic DNA is a prerequisite for successful PCR analysis. Time consumption, ease of operation and procedure cost are important secondary considerations for selecting an effective DNA extraction method. The simple, reliable and rapid DNA extraction method developed by Edwards and colleagues in 1991 [1] has proven to be the gold standard.ResultsThrough modification of the Edwards method of extraction, we have developed a one-tube protocol that greatly improves the efficiency of plant DNA extraction and reduces the potential for sample contamination while simultaneously yielding high quality DNA suitable for PCR analysis. We further show that DNA extracts prepared with this method are stable at room temperature for at least three months.ConclusionThe one-tube extraction method yields high quality plant DNA with improved efficiency while greatly minimizing the potential for cross contamination. This low-cost and environment-friendly method is widely applicable for plant molecular biology research.

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