qPCR

Southern Research offers a comprehensive range of quantitative PCR services to meet our clients' research needs. Specifically, our scientists have extensive experience in primer and probe design, PCR optimization, target sequence validation, and data analysis and interpretation.

qPCR Laboratory Capabilities

The qPCR laboratory at Southern Research provides services to determine the biodistribution of gene therapy products as well as the detection and quantitation of either DNA or RNA sequences in a variety of biological matrices. Moreover, all procedures are performed in compliance with U.S. Food and Drug Administration's (FDA) Good Laboratory Practice (GLP) regulations (21 CFR Part 58 and 21 CFR Part 11). Our laboratory maintains validated instruments and well-established standard operating procedures (SOPs), including: instrument operation and maintenance; sample collection; tissue lysation; DNA/RNA isolation; DNA/RNA quantitation; PCR setup; and data analysis. Since the qPCR laboratory began operation in 2001, many GLP studies have been completed covering a variety of DNA and/or RNA targets in different biological matrices.

Vector Biodistribution Studies

qPCR/RT-qPCR assays for vector copy number determination can be custom-designed and validated for multiple matrices and species. Southern Research has extensive experience analyzing adenovirus, herpes virus, retrovirus, and adeno-associated virus vectors for biodistribution in mice, rats, dogs, mini pigs, rabbits, and nonhuman primates.

Gene Expression Assays

We can provide gene expression assays for mouse and rat models as well as human samples. Specifically, multiple disease and cellular pathways are available, including: apoptosis; cancer; reproduction; toxicology and drug metabolism; and development and stem cells. Expression assays can be custom-designed for client-specified targets, or they can be ordered using Applied Biosystems' extensive TaqMan^® Expression Assays or Qiagen's QuantiTect Primer Assays. Our 384-well automated qPCR system is capable of analyzing more than 32,000 reactions per week, allowing researchers to collect a larger volume of data in a more time-efficient and cost-effective manner.

Viral/Bacterial Load Screening

Copy number as well as a plus/minus determination can be performed to determine the viral/bacterial load for a variety of matrices. We have experience in working with influenza, SARS, and anthrax in addition to many types of rodent pathogens.

miRNA Profiling and Quantitation

Micro-RNA assays for a variety of species can be used to analyze multiple matrices for miRNA expression profiles for distinct cellular pathways or for entire miRNomes.

Validation

Our GLP-compliant laboratory can assist in the design, development, and validation of custom qPCR assays to support preclinical GLP toxicology studies and human clinical trials. Alternatively, clients can use one of our pre-validated assays, including the adenovirus serotype 5 assay, to assist in reducing study costs for FDA-required gene therapy and vector-based therapeutic biodistribution testing.

Facility and Equipment

The qPCR laboratory features two instrument platforms that include an Applied Biosystems 7900HT that combines 96- and 384-well plate capability with fully automated robotic loading as well as two LightCyclers (Roche, Molecular Biochemicals, Mannheim, Germany). The 7900HT Quantitative PCR system, with interchangeable 96- and 384-well blocks and a hand-held integrated bar code reader, allows for simplified sample tracking and 24-hour automated plate handling for continuous, high-throughput data analysis.

The Roche systems are rapid microvolume thermal cyclers with built-in fluorometers. Fluorescence-labeled PCR products are monitored on line and in real time. The microvolume system allows for faster results than a Peltier-based system when smaller concentrations of DNA/RNA are to be analyzed.

The qPCR laboratory also utilizes an Agilent 2100 Bioanalyzer for RNA quality control after sample extraction. RNA samples are analyzed for their RNA integrity number (RIN) to ensure accurate data from downstream processing, particularly when gene or miRNA expression studies are being conducted. Additionally, our state-of-the-art laboratory facilities are designed with sample integrity in mind. Separate laboratories for sample receipt, preparation, and data analysis reduce the possibility of sample contamination. All PCR suites are BSL-2 certified and fully GLP-compliant.