UNLABELLED PREMISE OF THE STUDY Development of genetic markers can be costly and time-consuming, especially when multiple primer pairs are fluorescently labeled. This step was streamlined by combining two techniques in the same PCR reaction: (1) custom-labeling of primers by the investigator and (2) multiplexing multiple primers together in the same reaction. • METHODS AND RESULTS This tec...

Reproducible and target-specific polymerase chain reaction (PCR) amplification relies on several interrelated factors of which primer design is central. Here, we describe new free bioinformatics software, the FastPCR which was developed, and continues to be updated, based on detailed experimental studies of PCR efficiency for the optimal design of primers and probe sequences and for repeat sear...

A polygalacturonase-inhibiting protein (pgip) gene from Malus domestica cv Granny Smith apple fruit was cloned by degenerate oligo-primed polymerase chain reaction (PCR) and Inverse PCR. An alignment of the pear and bean PGIP sequences was used to design degenerate PCR primers in highly conserved regions. Degenerate PCR allowed the amplification of a 351bp internal fragment of the pgip gene, te...

OBJECTIVE To identify associations between microbes and host genes in cats with feline chronic gingivostomatitis (FCGS), a debilitating inflammatory oral mucosal disease no known cause, compared healthy periodontitis (control cats). ANIMALS 19 control 23 FCGS. PROCEDURES At least 1 caudal swab specimen was obtained from each cat. Each underwent unbiased metatranscriptomic next-generation RNA se...

Invasive aspergillosis (1 is a life-threatening condition in immunocompromised patients. An early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. Recently, the polymerase chain reaction (PCR) has been used successfully in detecting specific DNA of several pathogen. In this study, nested PCR was used to detect DNA specific for A!.pergiflus s...

PrimerStation (http://ps.cb.k.u-tokyo.ac.jp) is a web service that calculates primer sets guaranteeing high specificity against the entire human genome. To achieve high accuracy, we used the hybridization ratio of primers in liquid solution. Calculating the status of sequence hybridization in terms of the stringent hybridization ratio is computationally costly, and no web service checks the ent...