step. The presence of Vent DNA polymerase and PCR products other than the intended DNA linker, however, did not seem to interfere with subsequent reactions because ligations using both gel-purified and unpurified PCR products yielded a similar number of transformants. Eight transformants were tested, all of which contained the PCR product as an insert. The yield was considered to have high effi...

Polymerase chain reaction (PCR) with subsequent restriction fragment length polymorphism (RFLP) analysis is often used for phytoplasma identification and classification. Although these techniques are very sensitive and specific, in some cases, nonspecific reactions, false positives and negatives results, as well as unusual or illegible profiles after RFLP analyses, amplification of plant host ́s...

Screening of 1,750 pneumococcal isolates for common serotypes by PCR was followed by Quellung reaction analysis of PCR-negative isolates with a comparison to the conventional (Quellung reaction only) approach. PCR agreed with Quellung reaction results for 99% of isolates. The sequential PCR/Quellung reaction algorithm is accurate and more cost-effective than the conventional approach.

A real-time PCR was designed for detection of Chlamydophila pneumoniae and Mycoplasma pneumoniae such that each pathogen could be detected in a single tube and differentiated using molecular beacons marked with different fluorochromes. This duplex PCR, targeting the P1 adhesion gene for M. pneumoniae and the ompA gene for C. pneumoniae, was compared with two conventional PCR assays targeting th...

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 thermo...