Flow Cytometric Quantification of Competitive Reverse Transcription-PCR Products

Flow cytometric quantification of competitive reverse transcription-PCR products.

BACKGROUND Competitive PCR of reverse transcribed mRNA sequences is used to quantify transcripts, but the usual approaches are labor-intensive and time-consuming. We describe the non-gel-based quantification of competitive reverse transcription (RT)-PCR products with use of microparticles and flow cytometry. METHODS PCR products of a target sequence and an internal control sequence (competito...

Flow Cytometric Analysis of Reverse Transcription-PCR Products: Quantification of p21 and Proliferating Cell Nuclear Antigen mRNA

Background: Reverse transcription-PCR (RT-PCR) is a powerful tool in clinical diagnostics for analyzing even small amounts of RNA, but sensitive assays for quantifying the amplification products are time-consuming or expensive. Here we describe a novel flow cytometrybased assay for rapid and sensitive determination of relative amounts of RT-PCR products. Methods: For flow cytometric quantificat...

Quantitative competitive reverse transcription-PCR for quantification of dengue virus RNA.

A quantitative competitive reverse transcription-PCR assay was developed to quantify dengue virus RNA in this study. The main features include a primer pair targeting a highly conserved region in the capsid and the addition of competing RNA that contains an internal deletion to provide a stringent internal control for quantification. It can be utilized to quantify RNA isolated from the four den...

Quantification of Clostridium botulinum toxin gene expression by competitive reverse transcription-PCR.

Clostridium botulinum produces a characteristic botulinum neurotoxin which can cause an often fatal neuroparalytic condition known as botulism. Although food-borne botulism is rare, critical screening by food companies is necessary to ensure that food products are safe. At present, the food industry assesses the risks of botulinum neurotoxin production by challenge testing to check any new food...

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