High-Throughput Real Time PCR Core

Locations: Applied Biosystems, Foster City, CA

Overview | Investigators | News | Background | Resources

Overview

The ITN has established a partnership with Applied Biosytems to perform high-throughput multiplex real-time PCR on 1000 genes simultaneously. These target genes have been selected by ITN members and Celera genomics for their role in immune responses and/or tolerance induction. The complete list of these genes is publicly available in the Resources section of this website.

The protocol used by the Applied Biosystems core uses a unique pre-amplification step that has dramatically reduced the initial amount of RNA required for PCR. This new protocol consists of 10 PCR cycles using all ITN primer sets in a single reaction. "Kiloplexing" as it has been termed, has increased the sensitivity of the method from 50% detection to approximately 85%. The required sample size is reduced from 30 ug to only 700 ng using this technique.

The High-throughput Real Time PCR Core is complementary to the ITN's cDNA microarray facility and will assess gene expression changes in cases where RNA is quite limited, for example tissue biopsy material.

Participating Investigators

Mimi Roque-Biewer, Applied Biosystems

News & Recent Developments

	Pushing the limits of real time PCR: Collaboration with Applied Biosystems gets results - ITN eXpress [go]
	Friend and Foe: In Search of a More Tolerant Immune System - BioBeat [go]
	Applied Biosystems And Immune Tolerance Network Announce Immunology Research Agreement - Applera [go]

Background Articles

	Quantification of mRNA using real-time RT-PCR: trends and problems - J Mol Endocrinol [go]
	Real-time multiplex PCR assays - Methods [go]
	An overview of real-time quantitative PCR: applications to quantify cytokine gene expression - Methods [go]

Resources & Interesting Links

	Celera Genomics [go]
	Applied Biosystems [go]