Identification of Introns by Reverse-Transcription PCR

Identification of introns by reverse-transcription PCR.

analyzing the gene of interest has several advantages over subcloning analysis. This method does not require any information about physical maps of the insert DNA. Positions of transposon insertion are presented as a distance from one end of the genomic DNA. Even if the restriction map is known, this technique should be preferable because of its simplicity. Using this protocol, it takes only on...

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

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; ...

Identification of stress-responsive genes in Streptococcus mutans by differential display reverse transcription-PCR.

Streptococcus mutans, which causes dental caries in the human oral cavity and occasionally causes infective endocarditis in the heart, withstands adverse environmental stress through diverse alterations in protein synthesis. Differential gene expression in response to environmental stress was analyzed by RNA fingerprinting using arbitrarily primed PCR with a panel of 11mer primers designed for ...

Identification of a cytochrome P450 gene by reverse transcription--PCR using degenerate primers containing inosine.

A cytochrome P450-like gene, tentatively named P450CMEF, was amplified by a mixed oligonucleotide-primed amplification of cDNA from C3H mouse embryo fibroblast cells, designated 10T1/2, that had been treated with 7,12-dimethylbenz[a]anthracene (DMBA) or benz[a]anthracene (BA). A set of inosine-containing degenerate primers that were targeted to two conserved regions of known cytochrome P450 cDN...