Real-time Reverse Transcription Quantitative PCR

Overview

Real-time quantitative PCR (qPCR) is a highly sensitive and specific method for amplifying, detecting and quantitating target transcripts. The Gene Expression Core (GEC) provides qPCR services using the Applied Biosystems (ABI) 7900HT Sequence Detection System and the Fluidigm BioMark Dynamic Array.

For both platforms, there are options available for fluorescent (SYBR or EvaGreen) or TaqMan probe-based detection of mRNA and miRNA target molecules.

Details

  • Fluidigm BioMark Dynamic Array. The Fluidigm BioMark Dynamic Array provides high-throughput qPCR analyses using very little reagent master mix. Ten nanoliter reactions are conducted in array platforms for 48 samples x 48 targets, and 96 samples x 96 targets. Originally created to be used with the ABI TaqMan platform, the dynamic array can also be used in conjunction with an EvaGreen detection method.

    Standard first- and second-strand reverse transcription reactions are conducted with the starting RNA material. Subsequently, the cDNA is loaded into the dynamic array and preamplified with a gene-specific primer pool before undergoing a final amplification.

  • Applied Biosystems 7900HT Sequence Detection System. The GEC also provides standard qPCR on the ABI 7900HT Sequence Detection System in 96- and 384-well standard and FAST block formats. In addition, the GEC can perform expression analysis and target detection on the ABI TaqMan Low Density Array.

Fees

The cost for each project is dependent on experimental design, vendor costs and time required for completion. This information is provided in each project proposal, which must be approved by the principal investigator prior to project initiation.

Contact

For more information or to initiate a project, contact us at 507-255-1086 or by email.