The effects of 5 factors (template DNA, Mg(2+), dNTPs, Taq DNA polymerase, and primer) on the polymerase chain reaction (PCR) were investigated to optimize the start codon targeted polymor-phism (SCoT)-PCR system of Dactylis glomerata L., using an orthogo-nal design L16 (4(5)). A suitable SCoT-PCR system for D. glomerata was established; the 20 μL reaction volume contained 3.0 mM Mg(2+), 0.2 mM...

We report here the rational design and optimization of an antibody-responsive, DNA-based device that enables communication between pairs otherwise non-interacting proteins. The is designed to recognize bind a specific antibody and, in response, undergo conformational change leads release DNA strand, termed “translator,” regulates activity downstream target protein. As proof principle, we demons...

Point mutations in somatic cells play a role in the etiology of several classes of human pathologies. Experimental procedures are required that allow the detection and quantitation of such mutations in disease-related genes in tissue biopsy samples without the need for the selection of mutated cells. We describe the genotypic analysis of single base pair mutations in the Taq I endonuclease reco...

DNA analysis is a key procedure in genetic engineering. Nowadays the analysis is often done by PCR with Taq DNA polymerase. Although the last enzyme price is quite low, demand for numerous analyses results in much money expenditure which are not affordable for many laboratories. In a meanwhile, many screening tasks do not require the highly purified enzyme. Taking into account the enzyme unique...

Random amplified polymorphic DNA (RAPD) is a simple, fast and cost effective method for assessing genetic diversity of plant varieties. The reproducibility of the RAPD amplification was affected by several factors, therefore causing misinterpretation. In this study, we analyzed the effect of changing the concentration of magnesium chloride, template genomic DNA and Taq DNA polymerase with the o...

To evaluate PCR-generated artifacts (i.e., chimeras, mutations, and heteroduplexes) with the 16S ribosomal DNA (rDNA)-based cloning approach, a model community of four species was constructed from alpha, beta, and gamma subdivisions of the division Proteobacteria as well as gram-positive bacterium, all of which could be distinguished by HhaI restriction digestion patterns. The overall PCR artif...