Real-Time quantitative PCR (qPCR) is an accurate and sensitive method for quantifying the abundance of a target DNA sequence. This can be from genomic DNA or from cDNA resulting from the reverse-transcription of RNA. A polymerase chain reaction (PCR) is performed using sequence-specific primers and a DNA-binding dye or probe that provides a fluorescent signal proportional to the amount of product (or amplicon) formed during each amplification cycle.
Analysis of the amplification curves allows samples to be quantified via a standard curve, or used to calculate relative expression levels between samples (e.g. for validation of DNA arrays or confirming gene knockdown by siRNA).
The aim of the laboratory is to facilitate the effective use of the qPCR technique by the INP investigators. It is also open to others researchers present on the campus. Researchers of others campus and non-university users may also take advantage of qPCR Support Facility. For pricing information please contact the scientific coordinator.
The facility provides access to two qPCR apparatuses: one Applied Biosystems 7500 Fast and one Biorad CFX96 equipped with 96 well blocks. This platform support SybrGreen, Taqman, Molecular beacons or Scorpion reaction chemistries and is calibrated for the broadest range of dyes available: FAM™/SYBR® Green I, VIC®/JOE™, NED™/ TAMRA™/ Cy3™, ROX™/Texas Red®, and Cy5™ dyes. These apparatuses offer maximum performance in the minimum time (40 minutes).
The facility has also acquired an UV PCR cabinet with an HEPA filtration system for preventing contamination during the setting up of PCR reactions
The users must provide the supplies and reagents for the assays.