Improved quantitative real-time RT-PCR for expression profiling of individual cells

Improved quantitative real-time RT-PCR for expression profiling of individual cells.

The real-time quantitative polymerase chain reaction (rtqPCR) has overcome the limitations of conventional, time-consuming quantitative PCR strategies and is maturing into a routine tool to quantify gene expression levels, following reverse transcription (RT) of mRNA into complementary DNA (cDNA). Expression profiling with single-cell resolution is highly desirable, in particular for complex ti...

Improved quantitative real-time RT±PCR for expression pro®ling of individual cells

The real-time quantitative polymerase chain reaction (rtqPCR) has overcome the limitations of conventional, time-consuming quantitative PCR strategies and is maturing into a routine tool to quantify gene expression levels, following reverse transcription (RT) of mRNA into complementary DNA (cDNA). Expression pro®ling with single-cell resolution is highly desirable, in particular for complex tis...

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...

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