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PCR was performed in 20 l reaction mixture containing 5 ng of the obtained cDNA (or 50 ng of human genomic DNA as a control), 1x Phusion HF buffer, 0.2 mM dNTP mix, 0.5 M of each primer and 0.4 U Phusion DNA polymerase (F530S, Thermo Scientific)

To monitor the transcript level of the trxR gene, the hybridizing probe was amplified from D141 genomic DNA with the primer pair JB064/065 (5-TGTTCTCTATGAGGGTATGC-3 and 5-TTCGACTTGTATTCTGGAACG-3) and labeled during amplification by applying the PCR DIG labeling mix (Roche)

Studies examining 84 genes involved in neurotransmission showed: Changes in 45 gene expression levels one hour post-administration Sustained alterations in 22 genes at three hours High correlation between Selank peptide and GABA-induced expression changes Unique gene targets including dopamine receptors and GABA transporters These molecular findings provide mechanistic insights into Selanks rapid onset and sustained effects

Addition of free Fe 3+ to the reduced product brings about the formation of the intensive Perls Prussian blue complex, Fe 4 [Fe(CN ) 6 ] 3 , which has a strong absorbance at 700 nm