dna amplification using taq dna polymerase is one of the most widely used techniques in molecular biology and biotechnology. the aim of this study was to amplify the gene of this enzyme from a thermophilic bacteria called thermus aqauticus and clone it into a vector for future use. using specific primers the cdna of taq dna polymerase was amplified and ligated into the cloning vector ptz57r usi...

The Telomeric Repeat Amplification Protocol (TRAP) and its modified versions (including ours, TP-TRAP) change the size and/or the ratio of the telomerase products in the amplification stage of the assay. Based on our recently published method we developed a new TRAP. This method ensures that the number of telomeric repeats present in the original telomerase products does not change on PCR ampli...

The esterase-based insecticide resistance mechanisms characterised to date predominantly involve elevation of activity through gene amplification allowing increased levels of insecticide sequestration, or point mutations within the esterase structural genes which change their substrate specificity. The amplified esterases are subject to various types of gene regulation in different insect speci...

We describe a simple method for the cloning of PCR products without the need for post-amplification enzymatic treatment. Tailed PCR primer sets are used to create complementary staggered overhangs on both insert and vector by a post-PCR denaturation-hybridisation reaction. The single-stranded overhangs are designed to allow directional cloning in a ligase-free manner. This 'enzyme-free cloning'...

Enzyme amplification has proved to be a highly sensitive quantification technique for immunoassays. We have shown that by using a fluorescent end-point, even more sensitive enzyme amplification assays can be generated than hitherto reported. We describe some general properties of this system and demonstrate its application in an assay for human proinsulin in plasma. The detection system can be ...

Nanostructures Chengzhou Zhu,†,§ Guohai Yang,†,§ He Li,†,§ Dan Du,† and Yuehe Lin*,†,‡ †School of Mechanical and Materials Engineering, Washington State University, Pullman, Washington 99164, United States ‡Pacific Northwest National Laboratory, Richland, Washington 99352, United States ■ CONTENTS Nonenzymatic Sensors 231 Glucose 231 Hydrogen Peroxide 231 Cation 232 Anion 232 Other Species 232 ...

In plants, secondary metabolites and polysaccharides interfere with genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The removal of such contaminants needs complicated and time-consuming protocols. In this study, a simple, rapid and efficient method for leaf DNA extraction was optimized. This method use small amount of plant mater...

Genomic DNA extraction with a high quantity and quality is a basic requirement in molecular biology. The DNA obtained was free of any contamination proteins, polysaccharide, polyphenols and colored pigments. These compounds would interfere with the genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The isolated genomic DNA was fo...

Numerous high-throughput genomics assays require the amplification of a large number of genomic loci of interest. Amplification is cost-effectively achieved using several short single-stranded DNA sequences called primers and polymerase enzyme in a reaction called multiplex polymerase chain reaction (MP-PCR). Amplification of each locus requires that two of the primers bind to the forward and r...