Degenerate primers-based polymerase chain reaction (PCR) are commonly used for isolation of unidentified gene sequences in related organisms. For designing the degenerate primers, we propose the use of local alignment search method for searching the conserved regions long enough to design an acceptable primer pair. To test this method, a WD40 repeat-containing domain protein from Beauveria bass...

To increase detection sensitivity, we modified primers using complementary locked primer (CLP) technology. The sensitivity of the reverse transcription-PCR (RT-PCR) with CLP-modified primers was 10- to 100-fold higher than that of RT-PCR without these primers. CLP-modified primers can increase sensitivity, providing a widely accessible method for molecular diagnosis.

We evaluated the usefulness of PCR assays that target the internal transcribed spacer (ITS) region for identifying mycobacteria at the species level. The conservative and species-specific ITS sequences of 33 species of mycobacteria were analyzed in a multialignment analysis. One pair of panmycobacterial primers and seven pairs of mycobacterial species-specific primers were designed. All PCRs we...

Olive tree is one of the most important oleaginous crops in the world, and known for having large genetic variability. Application of molecular markers is a suitable tool to investigate the genetic pool in crops. Therefore, in the current study 29 olive cultivars were tested by ten primers constituted by inter simple sequence repeats (ISSR). Plant materials were obtained from the collection of ...

In Vibrio cholerae, the etiological agent of cholera, most of the virulence genes are located in two pathogenicity islands, named TCP (Toxin-Co-regulated Pilus) and CTX (Cholera ToXins). For each V. cholerae pathogenicity gene, we retrieved every primer published since 1990 and every known allele in order to perform a complete in silico survey and assess the quality of the PCR primers used for ...

background: splicing by overlap extension (soe) pcr is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function.objectives: we introduced a nested-soe-pcr (n –soe-pcr) in order to increase the specificity and generating mutations in a gene by soe-pcr.  materials and methods: genomic dna from bacillus thermoca...