About Real Time PCR
The Molecular Genomics Core (MGC) offers both custom and
pre-optimized assays for an assortment of applications. We offer
support in all aspects of the project from assay design to data
analysis. It’s our attention to detail from sample QC to statistical
analysis that sets our service apart from all others. Real Time PCR is
the gold standard for validating gene expression data. Typically a
list of genes with associated NCBI accession numbers are submitted for
design of either TaqMan or SYBR assays. Whenever possible, assays
will be designed such that a primer or TaqMan probe transverses an
exon-exon junction in order to reduce the contribution of genomic DNA.
The resulting probe/primers will be compared to the public database to
ensure specificity. Samples should be submitted in the form of total
RNA either frozen or as an ethanol precipitate. Once received, sample
quantity will be determined via the NanoDrop which will be used to
normalize RNA input for the reaction. Two hundred nanograms of sample
will then be used to assess the integrity of the sample via the Agilent
BioAnalyzer. Upon completion of QC, real-time analysis with be
carried out on our ABI 7500 and a detail report generated. Each report
consists of sample concentration, sample quality (Electropherogram and
Gel-like image), relative ratios, or absolute values.
Real-Time PCR Services
- Gene expression profiling
- siRNA knockdown validation
- Allelic discrimination / SNP genotyping
- Transgenic mouse genotyping
Service Features & Options
- Sample Quantitation via NanoDrop® ND-1000
- Qualitative Sample analysis via Agilent BioAnalyzer®
- Assay (primer/probe) design & optimization
- SYBR/TaqMan Chemistry Options
- Relative/Absolute Quantitation Options
- Easy to understand data report with statistical analysis
Useful Links
To schedule an appointment or to discuss experimental design, email Dr. Haiping Hao