Simon Baker PhD, Global R&D Director, Bioline Reagents Ltd: discusses MicroRNA Profiling Using a Rapid and Highly Sensitive qPCR Panel.
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Test a range of annealing temperatures. Depending on the qPCR results, the annealing temperature should be increased or decreased in 2-3oC increments. This can be done in a single experiment using a thermal gradient. Alternatively, a range of annealing temperatures should be tested using multiple qPCR experiments.
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