Real-time quantitative PCR analysis of human lung samples. Homo sapiens

Human lung samples from control and Down Syndrome prenatal patients were examined for Angiogenic Factors Overall design: qPCR gene expression profiling was performed to analyze the gene expression differences between human prenatal Down Syndrome patients and age matched control samples. Human lung tissue was obtained from the University of Maryland, Baltimore through its NICHD Brain and Tissue Bank for Developmental Disorders (NICH Contract #HHSN275200900011C, Ref. No. N01-HD-9-0011). The studies included in this report were examined by an institutional review board (IRB) of the University of Colorado School of Medicine and the Children’s Hospital Colorado and are supported through the Decedent Research Certification Program All tissues were collected, stored and distributed while maintaining strict confidentiality, meeting appropriate HIPPA standards, with oversight provided by the University of Colorado IRB. Accordingly, limited clinical data is available. Small numbers of tissue were available for study, including 5 DS and 4 control flash frozen samples for quantitative RT-PCR. Total RNA was extracted from human lung tissue using the RNAqueous Total RNA Isolation Kit (Life Technologies AM1912). Samples were DNase I treated with the Ambion TURBO DNA-free kit (Invitrogen AM1907) and tested for quality with an Agilent 2100 Bioanalyzer. Samples were reverse-transcribed with RT2 First Strand Kit (Qiagen 330401). These samples were amplified using the RT2 Profiler PCR Array Human Angiogenesis (Qiagen PAHS-024Z). Quantitative RT-PCR was performed on a LightCycler LC480 System (Roche).