Download SUPPLEMENTARY FIGURE LEGENDS Suppl Figure 1: Schematic

SUPPLEMENTARY FIGURE LEGENDS
Suppl Figure 1: Schematic representation of evaluated hybridization assay protocols.
a) Protocol A: biotinylation of multiplex PCR products was performed by using
biotinylated dCTP in an allele specific primer extension (ASPE) PCR,
b) Protocol B: cDNA was synthesized in a solid phase, and single-strand PCR was
performed using specific forward primers which contained a common extension in
their 5’-end,
c) Protocol C: All forward primers used in multiplex PCR were biotinylated at their
5’-end. All reverse primers had a specific extension, complementary to the couplingsequence of specific, commercially available, Luminex beads,
d) Protocol D: Forward primers for each gene, had a common extension, followed by
another specific sequence complementary to the coupling-sequence of specific,
commercially available, Luminex beads
Suppl. Figure 2: Liquid bead array assay optimizations. A) Multiplex PCR:
optimization of PCR cycle number, B) optimization of the number of beads, C)
optimization of hybridization temperature, D) Optimization of biotinylated PCR
product’s volume, E) effect of the size of PCR product in MFI and S/N