Real-Time PCR

  • The Real-Time PCR Laboratory is equipped with the 7900HT Fast Real-Time PCR system and the 7500 Fast Real-Time PCR system for detection and quantification of nucleic acid sequences. We support both SYBR Green dye and TaqMan Probe-based flourogenic 5’ nuclease chemistries. We have four different cycling blocks for the multipurpose 7900HT system, allowing for the following run modules:

    • 96-well Standard
    • 96-well Fast
    • 384-well Standard
    • 384-well TaqMan Low Density Arrays

    Our laboratory can perform both Real-Time PCR (quantification) and Endpoint (allelic discrimination, plus/minus) assays. For inclusive information about selecting the appropriate chemistry, assay type, and analysis method see Applied Biosystems Real-Time PCR Systems Chemistry Guide or contact us for consultation.

    Example applications include:

    • Detection of specific gene or allele
    • Quantitative gene expression (discover approximate copy number or compare expression levels in different types of samples)
    • Verification of microarray hits (see our Affymetrix and spotted array capabilities)

    We offer basic and full-service packages.

    Full-service 2-step Real-Time PCR assay*

    Customer Provides: Isolated and purified RNA template (contact us for preparation requirements) and custom primer/probe** set.

    Genomics Core Facility Provides:

    • Step 1: Standard RT reaction, using ABI High-Capacity cDNA Reverse Transcription kit, which utilizes random primers to generate single-stranded cDNA.
      • Human Control RNA and GAPDH primer/probe set is employed for each experiment
      • Quality/quantity check prior to second step assay
      • Storage/return of cDNA to investigator
    • Step 2: Genetic analysis on 7900HT Fast Real-Time PCR System
      • The details of this step depend upon the type of assay and chemistries used, and are determined during initial consultation with the investigator.

    *Variations of this package are acceptable (i.e. submitting cDNA samples). Please contact
    us to discuss project details.

    **Primer/assay design is available. Please contact us about this custom service.

    Basic service

    Customer Provides: Prepared sample plate ready for genetic analysis on the 7900HT system.

    Genomics Core Facility Provides:

    • Set-up of appropriate cycling block and background monitoring
    • Set-up of data collection software
    • General maintenance of equipment
    • Data transfer