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Then I ran some hot water, pressed play on my mix, and soaked

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Amplification of cDNA was run in a 25 L reaction volume that contained the following constituents: 2 L of the synthesized cDNA, 10 M of each of gene-specific primer pair (Table 2), 2 U/L Taq DNA polymerase in 10 mM buffer Tris-HCl, pH 8.8 (supplemented with 1.5 mM MgCl 2 , 50 mM KCl, 0.1% Triton X-100), 10 mM of each dNTPs and DEPC-H 2 O

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Fourth, the first visit of a new patient where labs are pending