Real-Time Quantitative RT-PCR: Design, Calculations, and Statistics

Real-time quantitative RT-PCR: design, calculations, and statistics.

A novel method for real time quantitative RT-PCR.

A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR) using real time detection and the 5' nuclease assay has been developed. Cystic fibrosis transmembrane transductance regulator (CFTR) target mRNA is reverse transcribed, amplified, detected, and quantitated in real time. A fluorogenic probe was designed to detect the CFTR amplicon. Relative increase in 6...

Heparinase treatment of RNA before quantitative real-time RT-PCR.

Vol. 35, No. 6 (2003) BioTechniques 1141 the ABI PRISM® 7000 and the TaqMan Universal PCR Master Mix protocols (Applied Biosystems). After discovering that the RNA from the thecal cells had vascular endothelial growth factor (VEGF) amplification and that RNA from granulosa cells had no VEGF amplification, the 18S rRNA amplification (TaqMan Pre-Developed Assay Reagents; Applied Biosystems), whic...

Q-Gene: Processing Quantitative Real-time RT-PCR Data

SUMMARY Q-Gene is an application for the processing of quantitative real-time RT-PCR data. It offers the user the possibility to freely choose between two principally different procedures to calculate normalized gene expressions as either means of Normalized Expressions or Mean Normalized Expressions. In this contribution it will be shown that the calculation of Mean Normalized Expressions has ...

REVIEW Quantitative real-time RT-PCR – a perspective

The real-time reverse transcription polymerase chain reaction (RT-PCR) uses fluorescent reporter molecules to monitor the production of amplification products during each cycle of the PCR reaction. This combines the nucleic acid amplification and detection steps into one homogeneous assay and obviates the need for gel electrophoresis to detect amplification products. Use of appropriate chemistr...