The role of 3' exonuclease excision in DNA polymerization was evaluated for primer extension using inert allele specific primers with exonuclease-digestible ddNMP at their 3' termini. Efficient primer extension was observed in amplicons where the inert allele specific primers and their corresponding templates were mismatched. However, no primer-extended products were yielded by matched amplicon...

SUMMARY We have developed U-PRIMER, a primer design program, to compute a minimal primer set (MPS) for any given set of DNA sequences. The U-PRIMER algorithm, which uses automatic variable fixing and automatic redundant constraint elimination to tackle the binary integer programming problem associated with the MPS selection problem. The program has been tested successfully with 32 adipocyte dev...

Current genome walking methods are cumbersome to perform and can result in non-specific products. Here, we demonstrate the use of partially overlapping primer-based PCR (POP-PCR), a direct genome walking technique for the isolation of unknown flanking regions. This method exploits the partially overlapping characteristic at the 3' ends of a set of POP primers (walking primers), which guarantees...

Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to i...

Here, we present a novel method for SNP genotyping based on protease-mediated allele-specific primer extension (PrASE), where the two allele-specific extension primers only differ in their 3'-positions. As reported previously [Ahmadian,A., Gharizadeh,B., O'Meara,D., Odeberg,J. and Lundeberg,J. (2001), Nucleic Acids Res., 29, e121], the kinetics of perfectly matched primer extension is faster th...

Background: Species identification in commercially processed food and feed products is one of the important issues. This study was conducted to develop a genetic method for the detection of pig and cattle species in processed food and feed products using newly designed species-specific primers targeting mitochondrial 12S rRNA gene fragments. Methods: Two sets of specific primers were designed ...

in present study, a sybr green based real time pcr assay was developed for specific detection and quantification of bacillus subtilis in dough used for bread making. new primer pairs were designed to amplify a 212 base pair fragment of the apre gene. specificity of these primer pairs was confirmed with conventional and real time pcr methods. standard curves constructed using the threshold cycle...

This study was carried out to determine the presence and frequency of Anaplasma ovis and Anaplasma marginale in sheep and dairy cattle in West-Azerbaijan province, Iran. A total number of 200 blood samples were randomly collected via the jugular vein from apparently healthy cattle (100) and sheep (100). The extracted DNA from blood cells was screened using genus-specific (Anaplasma...

Molecular genetics, and in particular the development of the polymerase chain reaction (PCR), has provided a way to distinguish these organisms. One primer pair specific for the genus Giardia is available (Mahbubani et al. 1991) as is a primer pair specific for G.intestinalis, (Mahbubani et al. 1992). During experiments in this laboratory, designed to compare strains and species of Giardia usin...