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YOUR DATA: ,,, Direct Real-time RT-PCR on Sympathetic Neuron Lysates,,, ,,, Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs

YOUR DATA:
Direct Real-time RT-PCR on Sympathetic Neuron Lysates

Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs



Direct Real-time RT-PCR on Sympathetic Neuron Lysates

Quantitative RT-PCR is the most common method used for measuring mRNA levels from small numbers of cells. This type of analysis can be difficult when cell samples are limiting. Ambion's Cells-to-cDNA II Kit (patent pending) overcomes this limitation by using an RT-PCR compatible cell lysis buffer, eliminating RNA isolation altogether. The kit produces cDNA from cultured mammalian cells in less than 2 hours. Here Drs Dziennis and Habecker use the Cells-to-cDNA II Kit to perform real-time RT-PCR on small numbers of primary sympathetic neurons.

Dziennis S and Habecker BA (2003) Cytokine suppression of dopamine beta hydroxylase by extracellular signal regulated kinase-dependent and -independent pathways. JBC 278(18):15897904.

As part of a more detailed study, Dziennis and Habecker used sympathetic neurons to investigate the cholinergic differentiation factor suppression of dopamine--hydroxylase (DBH), a norepinephrine synthetic enzyme. They measured DBH mRNA levels using real-time RT-PCR to determine if activation of a MAP kinase pathway was crucial for suppression of noradrenergic function.
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