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Supplemental Figure 1 A and B: Specificity validation of the PCR products Supplemental Figure 1A: Agarose gel electrophoresis of qPCR amplicons Gel strength: 4% NuSieve GTG (FMC Bioproducts, Rockland, ME) low melting temperature agarose

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  1. Supplemental Figure 1 A and B: Specificity validation of the PCR products Supplemental Figure 1A: Agarose gel electrophoresis of qPCR amplicons Gel strength: 4% NuSieve GTG (FMC Bioproducts, Rockland, ME) low melting temperature agarose 1x TBE buffer (90 mmol/L Tris base, 90 mmol/L boric acid, 2 mmol/L EDTA) Ethidium bromide staining: 0.5 µg/mL 10x loading buffer: 20% Ficoll, 100 mmol/L EDTA ph=8.0, 1% SDS, 0.25% bromophenol blue Biometra electrophoresis chamber (Goettingen, Germany) Gel: 9x6.5x0.5 cm; 10V/cm; 10 µL amplicon/0.5 µg DNA-sizer/lane M 1 2 3 4 5 6 M bp -- 1353/1078/872 -- 603 -- 310 -- 281/271 -- 234 -- 194 -- 118 -- 72 Lanes and target sizes: M= peqGold DNA-Sizer XIII (peqlab, Erlangen, Germany) 1= TBP (227 bp) 2= CDH16 (72 bp) 3= PPIA (121 bp) 4= HPRT1 (127 bp) 5= HIF1A (90 bp) 6= PSA=KLK3 (154 bp) bp= base pairs

  2. Supplemental Figure 1B: Melting peak chart from a melting temperature (Tm) calling analysis of the PPIA RT-PCR amplicon with SybrGreen I detection on the LightCycler 480 instrument. The PCR product has a Tm of 81.3°C. Melting temperature program

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