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In order to suppress the nucleophilic nature of the guanidine group in arginine, it was deactivated as the monohydrochloride salt over the course of 4 steps, and then as trifluoroacetate for another 2 steps

The use of real-time PCR increased dramatically from 2000, in part because it offered greater sensitivity and higher throughput (Figure 24) (Deepak, Kottapalli, Rakwal)