Quantitation of mRNA by polymerase chain reaction. Nonradioactive PCR methods

Quantitation of mRNA by the polymerase chain reaction.

A method for the quantitation of specific mRNA species by the polymerase chain reaction (PCR) has been developed by using a synthetic RNA as an internal standard. The specific target mRNA and the internal standard are coamplified in one reaction in which the same primers are used. The amount of mRNA is then quantitated by extrapolating against the standard curve generated with the internal stan...

Nonradioactive quantification of mdr1 mRNA by polymerase chain reaction amplification coupled with HPLC.

We describe a new strategy for quantifying mRNA by using polymerase chain reaction (PCR) coupled with HPLC. PCR-amplified products are directly analyzed with a specific HPLC column and are quantified by standardization against a housekeeping gene, beta-actin. To evaluate the experimental conditions, we examined the multidrug-resistance-associated mdr1 gene expression in two drug-sensitive cell ...

Polymerase Chain Reaction (PCR)

The polymerase chain reaction (PCR) is a powerful and widely used technique that has greatly advanced our ability to analyze genes. Genomic deoxyribonucleic acid (DNA) present in cells contains many thousands of genes. This makes it difficult to isolate and analyze any individual gene. PCR allows specific DNA sequences, usually corresponding to genes or parts of genes, to be copied from genomic...

Quantitation of enteroviral RNA by competitive polymerase chain reaction.

The polymerase chain reaction (PCR) is a new diagnostic technique for the detection of enteroviral infection; however, it currently provides only qualitative results. The aim of this study was to adapt PCR for the accurate quantitation of enteroviral RNA in clinical specimens. For this purpose, we designed a standard RNA which was homologous to sequences at the 5' end of the coxsackie B3 entero...

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*