Simple DNA extraction method for SSR-PCR analysis from different tissues ofTapiscia sinensis, an endangered plant species

2015 ◽  
Vol 75 (3) ◽  
pp. 393 ◽  
Author(s):  
Xiao-Jun Zhou ◽  
Rong-Shan Yan ◽  
Ya-Nan Xu ◽  
Da-Wei Zou ◽  
Peng Zhao ◽  
...  
2008 ◽  
Vol 71 (12) ◽  
pp. 2497-2503 ◽  
Author(s):  
BEATRIZ SÁNCHEZ ◽  
MAR RODRÍGUEZ ◽  
EVA M. CASADO ◽  
ALBERTO MARTÍN ◽  
JUAN J. CÓRDOBA

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.


2010 ◽  
Vol 53 (2) ◽  
pp. 249-252 ◽  
Author(s):  
Nadja Santos Vitória ◽  
José Luiz Bezerra ◽  
Karina Peres Gramacho

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.


Plant Methods ◽  
2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Santiago Vilanova ◽  
David Alonso ◽  
Pietro Gramazio ◽  
Mariola Plazas ◽  
Edgar García-Fortea ◽  
...  

2020 ◽  
Author(s):  
Wei Hu ◽  
J. Clark Lagarias

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.


Author(s):  
Parthadeb Ghosh ◽  
Parthadeb Ghosh ◽  
Sanjib Kumar Chattopadhyay ◽  
Sinchan Adhikari ◽  
Soumen Saha ◽  
...  

2017 ◽  
Vol 9 (2) ◽  
pp. 866-870
Author(s):  
Sweta Sinha ◽  
Amarendra Kumar

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.


2016 ◽  
Vol 5 (08) ◽  
pp. 4754
Author(s):  
Tanushree Mitra* ◽  
Shivshankar Kumdale ◽  
Sameer Chowdhary ◽  
Amol D. Raut

The main objective of this study was to make sure whether randomly taken 12 samples were sensitive to abacavir. The genomic DNA from 12 blood sample were extracted by phenol chloroform DNA extraction method, extracted genomic DNA were amplified and sequenced, thereafter SNPs were detected. Every sample had shown the presence of normal base at SNP position. This study indicated, those randomly taken 12 patients were sensitive to abacavir, so they can consume abacavir if they get infected with HIV.


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