The polymerase chain reaction (PCR) is an indispensable DNA amplification technique that has been exploited in numerous areas, including molecular diagnostics. One major drawback of PCR is the competing amplification of undesired off-target products, which primarily occurs at ambient temperatures prior to PCR cycling (1). Hot Start PCR techniques aim to block the extension of primers in nonspec...

To make PCR a specific, efficient and cost effective tool for researchers and clinicians the most important aspect is oligonucleotide primer design. This review discusses various aspects of primer design. Advice is provided for optimal design and the role of bioinformatic tools is highlighted. The authors discuss theoretical considerations and compare computational and experimental studies.

Various biological and clinical applications require the quantification of the messenger RNA (mRNA) abundance of the biological materials under investigation. Reverse transcription-polymerase chain reaction (RT-PCR) is a popular technique for mRNA quantification. However, in these applications, multiple quantifications of mRNA are required from the same sample. Multiplex RT-PCR uses multiple pr...

Potato virus Y biberde enfeksiyon oluşturan en yaygın virüslerden bir tanesidir. Bu çalışmada, 23 biber çeşidi ve 30 ıslah hattının PVY-0 PVY-1 patotiplerine karşı reaksiyonları belirlenmiş pvr2 lokusundaki allel genleri mekanik inokulasyon moleküler yöntemler ile belirlenmeye çalışılmıştır. Her çeşit hatta ait dörder bitki, patotipleri ayrı inokule edilmiştir. İnokule edilen bitkiler, virüs si...

We have developed a primer-mediated PCR mutagenesis-based method for the generation of positive controls to test the sensitivity of single-strand conformation polymorphism (SSCP) or any other PCR-based mutation screening method. This technique is based on the incorporation of a third longer primer, containing a mismatched base, into the PCR along with the two wild-type primers normally used to ...

We developed a mutation-screening protocol for the ASS1 gene in order to guide clinical management of neonates with elevated citrulline detected during routine newborn screening. An exon-based amplification and sequencing method was designed and successfully applied to patients to identify disease-associated mutations. The sequencing-based method was applied to three patients with mild or asymp...