The process of insulating nucleic acid that is fast, simple, and high results is very important for high-quality DNA analysis. The ability of the Microgem Pdqex Phytogem System and Omega Bio-Tek EZNA® Kit Mini to extract DNA Ready PCR evaluated by extracting forensically relevant “legal” plant species: Ipomoea Purpurea, Artemisia Absinthium, Mitragyna Speciona, Datura Stramonium, and Papavon Somniferum.
The plant material was destroyed, processed using the manufacturer’s factory protocol for pdqex nucleic acid extraction or manufacturer’s protocol for omega extraction, quantified using fluorometer fluorometer 2.0 invitrogen, and analyzed for amplifiability with PCR which uses q rotor-gen qiagen instruments and published tests. DNA amplicons for legal high species produces a high resolution melting curve by melting observed when DNA isolated using Qiagen Dneeny Plant Mini Kit in previous research.
Simple installation of high-molecular molecular DNA extraction method that is suitable for single molecular technology.
High molecular weight and pure DNA are very important for high-quality results from the 3rd generation DNA analysis and optical mapping technology. Conventional core insulation methods to prepare DNA high molecular genomes from plant tissue include preparation of protoplasts or instill nucleus in the agarose matrix with further manipulation through electro-elution or gel electrophoresis of pulsating fields. In this method, this nuclei is isolated by a physical grinding network and rearranged the intact nucleus in the buffer of a unique nuclear insulation (NIB).
DNA plastid is released from organelles and removed with osmotic buffers by washing and centrifugation. The purified nuclei is then licis and further cleansed by organic extraction, and DNA genomes are deposited with high CTAB concentrations. GDNA is a very pure molecular weight, high extracted from nuclei, dissolved in a high pH buffer, allowing stable long-term storage.
This method is unique and avoids the use of embedding in agarose, which dramatically reduces the time (4-8) h versus days), complexity, and material costs. This procedure can basically be used in every plant species and network stage. Here we describe a case study and a simple method to quickly prepare high molecular weight gdna of highland cotton, blackgrass, and strawberries that are suitable for single molecular sequencing.
DNA extraction from leaf plants uses microneedle patches.
High quality DNA isolation from infected plant specimens is an important step for molecular detection of plant pathogens. However, DNA isolation from plant cells surrounded by rigid polysaccharide cells involves complicated steps and requires benchtop laboratory equipment. As a result, the extraction of plant DNA is currently locked up in a complete laboratory and sample preparation has become one of the main obstacles for molecular detection in the location of the plant pathogen.
To overcome this obstacle, the simple DNA extraction method of plant leaves has been developed. Microneedle (MN) patch made of polyvinyl alcohol (PVA) can isolate plants or pathogenous DNA from different plant species in one minute. During the DNA extraction, the Polymer Patch MN penetrated into the plant leaves and broke the walls of rigid plant cells to isolate intracellular DNA. The extracted DNA is the polymerase chain reaction (PCR) which is well-out without additional purification.
This minimum invasive method has been successfully extracted by Phytophot Infests DNA from infected tomato leaves. In addition, MN fillings can be used to isolate DNA from other plant pathogens directly in the field. Thus, it has great potential to be a fast sample preparation technique, in place for plant pathogen detection. © 2020 by John Wiley & Sons, Inc. Basic Protocol Based Patch-Based Patch 1: Microneenedle Patch Protocol Fabrication Support 2: Real-time PCR amplification of Microneedle Patch DNA extracted.
Extraction of non-destructive DNA from Herbarium specimens: a method that is very suitable for plants with small and fragile leaves.
The protocol for DNA extraction from plants generally involves physical damage and network chemistry. The use of this conventional method is blocking the preservation of morphological information from herbarium specimens, especially for small plants with several leaves, and reduces the value of the specimen voucher.
Here, we developed a new DNA extraction protocol, non-destructive (protocol 1) which only requires a small piece of leaf (<25 mm2) to get a suitable DNA for the DNA sequencing of fragile herbarium specimens. The protocol is very simple and fast; Extraction buffers are placed on the surface of the specimen leaf intact for 30 minutes at room temperature (20 ° C). The extracted DNA quality was examined by PCR amplification from two barcode DNA plastal barcode, Maturase K Gen (MATK, CA. 850 BP) and ribulose-1.5-bisphoschosboxy-1.5-oxygenase (RBCL, CA. 550 BP), for 14 species Vascular plants cover various taxonomy groups.
The protocol took the order of 80.0% specimens for MATK and 46.2% specimens for RBCL. Placing the extraction buffer to the specimen does not cause tears or deformation, but causes color changes in some plants. To improve DNA results for specimens that are not compatible with protocol 1, we develop alternative protocols for DNA extraction with the destruction of specimens (protocol 2) which is minimal invasive.
In this protocol, the leaf is soaked in the extraction buffer for 30 minutes and kept next in the fragment bag on the specimen sheet. This alternative method takes the sequence of MATK from 80.0% specimens and RBCL sequences of 92.8% of specimens.
The combination of protocol 1 and 2 allows us to get the MATK sequence of 90.0% specimens and RBCL sequences formed 92.8% of specimens. The new protocol facilitates the use of museum specimens for the use of DNA museum specimens while still preserving morphological information.