Simple method, low cost and safe use 96-Well round plate based on homemade filter paper and homemade solutions developed for high plant DNA extraction to be used in marker molecular analysis. Low-cost and safe methods are developed for high throughput extraction of plant DNA for molecular analysis. In this method, we describe a simple way to prepare 96 well spin plate using filter paper, plant material products for binding DNA.
The filter paper-based rotary plate can be combined with homemade non-toxic buffers for high throughput extraction from plant DNA. We confirm that filter paper is a solid phase DNA fastening material that is efficient and is proportional to silicon-based glass fiber filters adopted in the commercial DNA extraction kit, and that the DNA of the plant extracted with this method can be easily used as a template for PCR.
The efficacy of this method is also indicated fully by the analysis of marker molecules in separating tomato populations. Because costs are very reduced compared to commercial kits, this method has great value for small laboratories with limited resources.
DNA extraction method directly from parasitic fungi requires from infected plant tissue.
Flour mushrooms and rust mushrooms are compulsory parasites that cannot live without home organisms. They are difficult for culture in synthetic media in the laboratory. The extraction of DNA genomes is one of the basic molecular techniques used to study the genetic structure of the population.
In this study, 2 different DNA extraction methods, Chelex-100 and Cetiltrimethylamamium Bromide (CTAB), are used to extract DNA from Eumymus flour mushrooms and puccinia strisorformis f. Tr tritici. The polymerase chain reaction is carried out with RDNA-Special-Special RDNA-internal spacer sequence transcripts. Both DNA extraction methods are compared and analyzed.
The results showed that Chelex-100 and CTAB were effective for extracting DNA genomes from infected plant tissue. However, fewer DNA is needed for the Chelex-100 method than for the CTAB method, and the Chelex-100 method involves a little step, simpler and safer, and does not require organic solvents compared to the CTAB method.
DNA quality is evaluated with a chain reaction polymerase, and the results indicate that the DNA genome is extracted using the Chelex-100 method is better than using the CTAB method, and enough to study the genetic structure of the population.
Short communication methods are efficient for simultaneous RNA extraction and high-quality DNA from various plant tissue.
Determination of gene expression is an important tool for studying the biological process and depending on the quality of RNA extracted. Changes in gene expression profiles can connect directly with mutations in the order or change of regulation DNA in cytosine DNA methylation, which is an epigenetic sign.
The correlation of gene expression with the order of DNA or polymorphism of epigenetic signs is often desirable; For this, a strong protocol to isolate RNA and high-quality DNA simultaneously from the same sample is needed. Even though commercial kits and protocols are available, they are mainly optimized for animal networks and, in general, limited to RNA or DNA extraction, not both. In this study, we explain efficient and accessible methods to extract RNA and DNA simultaneously from the same sample of various plant tissue, using a little initial material.
Efficient protocols in high quality nucleic acid extraction from several arabidopsis thalianly network (for example, leaves, inflorescence rods, flowers, fruit, kotedon, seeds, roots, and embryos) and from other non-model plants, such as Avicennia Schaueriana (Acanthaceae) , Theobroma Cacao (Malvaceae), Pasphalum Notatum (Poaceae), and Sorghum Bicolor (poaceae).
The nucleic acid obtained is used as a template for downstream analysis, such as MRNA sequencing, a quantitative real-polymerase real-polymerase chain reaction, bisulfite treatment, and others; The result is comparable to those obtained with a commercial kit. We believe that this protocol can be applied to various plant species, helping to avoid technical bias and sampling, and facilitate several RNA and DNA dependent analysis.
DNA extraction from plant food supplements: the effect of various pharmaceutical expriners.
Consumption of plant food supplements (PFS) has grown globally, with increased misleading labeling and fraud practices also reported. Recently, the use of molecular biology techniques has been proposed to detect botanical counterfeiting, one possible fraud in PFS. However, the difficulty in recovering DNA from several PFS samples was explained. Aiming to use DNA-based methods for assertive identification of plant species in PFS, adequate DNA isolation is needed.
However, PFS often contains pharmaceutical excipients known to have adsorbent properties that can interfere with DNA extraction. Thus, the purpose of this work is to assess the effects of various excipients (powder, silica, iron oxide and titanium dioxide) on DNA recovery / amplification. For that purpose, the number of Corn DNA templates is known to be pHS or to model the mixture of excipients and be quantified by real-time PCRs.
The tested expenses prove a clear adsorption phenomenon that justifies the stunning effect on DNA extraction from PFS. The use of 10% Talc or 0.5% of the DNA adsorbed dye, produces negative PCR amplification.
For the first time, pharmaceutical excipients were proven to affect DNA extraction which described the inability to recover DNA from several PFS samples in previous studies.