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 thermoc...

We introduce quantitative polymerase chain reaction (qPCR) primers and multiplex end-point PCR primers modified by the addition of a single ortho-Twisted Intercalating Nucleic Acid (o-TINA) molecule at the 5'-end. In qPCR, the 5'-o-TINA modified primers allow for a qPCR efficiency of 100% at significantly stressed reaction conditions, increasing the robustness of qPCR assays compared to unmodif...

The polymerase chain reaction using only a single 'consensus' tRNA gene primer, or a pair of primers facing outward from tRNA genes, amplifies a set of DNA fragments in bacterial, plant and animal genomic DNAs. Presumably, these PCR fingerprints are mainly derived from the regions between closely linked tRNA genes. The pattern of the PCR products is determined by which genomes and which primer(...

A simple mass spectrometric based assay, the PinPoint assay, has previously been described for typing single nucleotide polymorphisms. The identity of a polymorphism is determined by mass differences of single base extended genotyping primers as determined by MALDI-TOF mass spectrometry. A simple method for multiplexing the assay is described, employing multiple primers with 5'oligo(dT) sequenc...

OBJECTIVES To develop and apply the polymerase chain reaction with confronting two-pair primers (PCR-CTPP) for detection and identification of hemoglobin E (Hb E). MATERIAL AND METHOD Fifty unrelated northern Thais were included in the present study. DNA was extracted from peripheral blood mononuclear cells and targeted to amplify by PCR-CTPP. The amplified product was analyzed and compared w...

The box jellyfish Chironex yamaguchii is a highly venomous cubomedusa, also known as “habu-kurage” in Japan. It significant threat to marine activities Okinawa southern Annually, envenomation of the species causes nearly 100 200 injuries Prefecture. present study developed species-specific real-time PCR assay analyze environmental DNA (eDNA) for rapid detection water samples. Primers and probe ...

We read with great interest the article by Mendez et al. (2) published in a recent issue evaluating different PCR primers for cytomegalovirus (CMV) detection. As CMV PCR becomes a frequently used assay in the management of immunocompromised patients, such as transplant recipients and human immunodeficiency virus-infected individuals, an increasing number of laboratories are establishing in-hous...

1.Blaseio, U. and F. Pfeiffer. 1990. Transformation of Halobacterium halobium: development of vectors and investigation of gas vesicle synthesis. Proc. Natl. Acad. Sci. USA 87:67726776. 2.Cline, W.S., W.L. Lam, R.L. Charlebois, L.C. Schalkwyk, and W.F. Doolittle. 1989. Transformation methods for halophilic archaebacteria. Can. J. Microbiol. 35:148-152. 3.Cohen, S.N., A.C.Y. Chang, and L. Hsu. 1...