Introduction of multiple mutations can be accomplished through phage M13-based site-directed mutagenesis using several oligonucleotides (6). The major drawback of this method is the low efficiency of generating all the intended mutations simultaneously (7,9). The alternative methods for introducing multisite mutations are based on polymerase chain reaction (PCR) (1) and ligase chain reaction (L...

Environmental sensing in bacteria often involves the concerted action of sensor kinases and response regulators. Degenerate oligonucleotide primers were designed on the basis of amino acid similarity in the response regulators of these two-component systems. The primers were used in PCR to specifically amplify an internal DNA segment corresponding to the receiver module domain from genes encodi...

SUMMARY We developed a program named MFEprimer for evaluating the specificity of PCR primers based on multiple factors, including sequence similarity, stability at the 3'-end of the primer, melting temperature, GC content and number of binding sites between the primer and DNA templates. MFEprimer can help the user to select more suitable primers before running either standard or multiplex PCR r...

A substantial percentage of the expense in constructing full-genome spotted microarrays comes from the cost of synthesizing the PCR primers to amplify the desired DNA. We propose a computationally-based method to substantially reduce this cost. Historically, PCR primers are designed so that each primer occurs uniquely in the genome. This condition is unnecessarily strong for selective amplifica...

It has been demonstrated that the use of additional information such as ESTs and protein homology can significantly improve accuracy of gene prediction. However, many sources of external information are still being omitted from consideration. Here, we investigate the use of product lengths from RT-PCR experiments in gene finding. We present hardness results and practical algorithms for several ...

We presented a novel platform for an ultrafast PCR system, called the Rotary PCR Genetic Analyzer, which incorporates a thermal block and resistive temperature detector (RTD) for thermal cycling control, a disposable PCR microchip, and a stepper motor. The influenza viral RNAs from H3N2, H5N1, and H1N1 were simultaneously identified with high sensitivity and speed.

Economic profit-driven food adulteration has become widespread in the dairy industry. One of most common forms is substitution low-priced milk for high-priced milk. This prompted regulatory authorities to focus on various means authenticity testing. So far, many methods have been developed. Since upgraded, which forced testing meet needs detection, include DNA-based PCR methods. and PCR-derived...

multiple sclerosis (ms) is characterized by multiple areas of inflammation, demyelination and neurodegeneration. infiltrating th1 cd4+ t cells secrete proinflammatory cytokines. they stimulate the release of some cytokines, expression of adhesion molecules and these cytokines may cause damage to the myelin sheath and axons. in this study, we analyzed plasma levels and gene expressions of five i...