Competitive RNA Templates for Detection and Quantitation of Growth Factors, Cytokines, Extracellular Matrix Components and Matrix Metalloproteinases by RT-PCR

Competitive RNA Templates for Detection and Quantitation of Growth Factors, Cytokines, Extracellular Matrix Components and Matrix Metalloproteinases by RT-PCR

Blog Article

Detection of low-abundance mRNAs by reverse transcription-polymerase chain reaction (RT-PCR) has become a standard technique to determine gene expression by tissues and cells in culture.The ability to determine relative or absolute copy number of specific mRNAs has been difJicult due to inadequate internal standards to control for sample-to-sample DETANGLER variation.The use of a synthetic RNA standard with identical sequences to the PCR primers allows reproducible quantitation between samples and assays.By designing multi-sequence templates, several specijic mRNAs can be quantitated using a single template.

Addition of multiple templates to a single RT reaction allows the quantitation of a large number of targetsffom as little as 4 pg of total RNA.In this report, we present a series of seven primer/template systems to detect and quantitate 52 specific messages, including 26 growth factors Collections and receptors, 8 extracellular matrix components, 10 matrix-rnodifying enzymes and their inhibitors and 8 cytokines.