Real Time PCR

Equipment/facility: Equipment

  • Location

    United States

Description

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

Fingerprint

Real-Time Polymerase Chain Reaction
Exons
RNA
Single Nucleotide Polymorphism
Ethanol
Gels
Databases
Gene Expression
DNA
Genes