Quantitative Assessment of Human β-Globin Gene Expression In Vitro by TaqMan Real-Time Reverse Transcription-PCR: Comparison with Competitive Reverse Transcription-PCR and Application to Mutations or Deletions in Noncoding Regions

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; ...

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...

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

Single-cell gene expression profiling using reverse transcription quantitative real-time PCR.

Even in an apparently homogeneous population of cells there are considerable differences between individual cells. A response to a stimulus of a cell population or tissue may be consistent and gradual while the single-cell response might be binary and apparently irregular. The origin of this variability may be preprogrammed or stochastic and a study of this phenomenon will require quantitative ...

In vivo analysis of Aspergillus fumigatus developmental gene expression determined by real-time reverse transcription-PCR.

Very little is known about the developmental stages of Aspergillus fumigatus during invasive aspergillosis. We performed real-time reverse transcription-PCR analysis on lung samples from mice with invasive pulmonary aspergillosis to determine the expression of A. fumigatus genes that are expressed at specific stages of development. In established infection, A. fumigatus exhibited mRNA expressio...