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Then, a mixture with the following components was added: 4 L of the GoScript TM 5 concentrated reaction buffer (Promega, Madison, WI, USA), 3 L of MgCl 2 (final concentration 1.55 mM, Promega, Madison, WI, USA), 1 L of deoxyribonucleotide triphosphates (dNTPs, 10 mM, Thermo Fisher Scientific, Waltham, MA, USA), 1 L of Rnase inhibitor (20 U/L, Thermo Fisher Scientific, Waltham, MA, USA), and 1 L of GoScript TM reverse transcriptase (Promega, Madison, WI, USA) (160 U/L), giving a total volume of 20 L