A comparative study of commercial real-time reverse transcription PCR kits for forensic body fluid identification

Real-Time Reverse Transcription PCR

Real-time, fluorescence-based reverse transcription polymerase chain reaction (RT-PCR) has been transformed from an experimental technology into a mainstream scientific tool for the detection of RNA. This is because of several factors: 1) it is a homogeneous assay, which eliminates the requirement for post-PCR processing; 2) it has a wide dynamic range; 3) there is little interassay variation; ...

Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results

Real-time reverse transcription polymerase chain reaction (PCR) is the gold standard for expression analysis. Designed to improve reproducibility and sensitivity, commercial kits are commonly used for the critical step of cDNA synthesis. The present study was designed to determine the impact of these kits. mRNA from mouse brains were pooled to create serial dilutions ranging from 0.0625 μg to 2...

mRNA Profiling: Body Fluid Identification Using Multiplex Real-Time PCR

High-throughput real-time quantitative reverse transcription PCR.

Extensive detail on the application of the real-time quantitative polymerase chain reaction (QPCR) for the analysis of gene expression is provided in this unit. The protocols are designed for high-throughput, 384-well-format instruments, such as the Applied Biosystems 7900HT, but may be modified to suit any real-time PCR instrument. QPCR primer and probe design and validation are discussed, and...

Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR

BACKGROUND During the 2015 outbreak of Middle East Respiratory Syndrome coronavirus (MERS-CoV), six different commercial MERS-CoV RNA detection kits based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) were available in Korea. We performed analytical and clinical validations of these kits. METHODS PowerChek (Kogene Biotech, Korea), DiaPlexQ (SolGent, Korea), Anyplex (S...