Multiplex PCR: Critical Parameters and Step-by-Step Protocol

Multiplex PCR: critical parameters and step-by-step protocol.

By simultaneously amplifying more than one locus in the same reaction, multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory. While numerous papers and manuals discuss in detail conditions influencing the quality of PCR in general, relatively little has been published about the important experimental factors and the common difficulties...

Single-Step Multiplex PCR Assay for Determining 92 Pneumococcal Serotypes.

For pneumococcal disease surveillance, simple and cost-effective methods capable of determining all serotypes are needed. Combining a single-tube multiplex PCR with fluorescently labeled primers followed by amplicon analysis using automated fluorescent capillary electrophoresis, each serotype of 92 reference isolates and 297 recently collected clinical isolates was successfully determined.

Simultaneous detection of three pome fruit tree viruses by one-step multiplex quantitative RT-PCR

A one-step multiplex real-time reverse transcription polymerase chain reaction (RT-qPCR) based on TaqMan probes was developed for the simultaneous detection of Apple mosaic virus (ApMV), Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) in total RNA of pome trees extracted with a CTAB method. The sensitivity of the method was established using in vitro synthesized viral trans...

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*

Step – By – Step

Approaches to formalisation of medical guidelines can be divided into model–centric and document–centric. While model–centric approaches dominate in the development of clinical decision support applications, document–centric, mark–up–based formalisation is suitable for application tasks requiring the 'literal' content of the document to be transferred into the formal model. Examples of such tas...