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350 Community Drive Manhasset, NY 11030Quantitative PCR (QPCR) or real-time RT-PCR is a sensitive, reproducible and accurate method for determining the DNA or mRNA levels in tissues or cells. The amount of PCR product produced in every cycle step of the PCR reaction is monitored, which may be accomplished directly with intercalating dyes which bind to the double stranded DNA (e.g. SYBR Green); or indirectly with fluorescent probes (e.g. TaqMan or Universal ProbeLibrary, UPL). In a conventional PCR reaction, the PCR product amount is measured as an end-point analysis on an agarose gel. In a QPCR reaction, the key feature is the ability to monitor the DNA amplification process as it happens, in “real-time.” QPCR delivers powerful capabilities for generating and analyzing data for a wide variety of applications. The Feinstein Institute’s QPCR Core Facility, located on the first floor of the North Wing in Rooms 1261 and 1263. houses 3 machines: one Applied Biosystems (ABI) 7900HT Real-Time PCR System with Sequence Detection System (SDS) software and two Roche Applied Science LightCycler480 PCR Systems with software. Each system is equipped to perform QPCR in a 96 or 384-well format. The ABI 7900HT is also equipped with a TaqMan Low Density Array (TLDA) block, for use with pre-loaded TaqMan Gene Expression Assays. The TaqMan Array Card is a 384-well micro fluidic card that enables a researcher to perform 384 simultaneous reactions without the need to use liquid-handling robots or multichannel pipettes to fill the card. This micro fluidic card allows for 1-8 samples to be run in parallel against 12-384 TaqMan Gene Expression Assay targets that are pre-loaded into each of the wells on the card. Researchers are able to use Applied Biosystems premade TLDA Gene Signature Panels (for Angiogenetics, Apoptosis, Alzheimer’s, Immune etc.) or design their own custom TLDA cards.
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