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Invitrogen/Thermo Fisher Scientific) was employed to generate cDNA and to carry a linear co-amplification (1114 cycles) of three amplicons: two derived from target genes (final concentration of each primer was 80 nM) and one derived from the RPL8 reference gene (final concentration of each primer was 60 nM)

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Like all proteins, FBP is heat-sensitive, and denaturation affects its folate binding capacity

Remove any air bubbles by tapping the syringe and gently pushing the plunger until a tiny droplet appears at the needle tip