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Genotyping

KT Kinga Tibori
VZ Veronika Zámbó
GO Gabriella Orosz
PS Péter Szelényi
FS Farkas Sarnyai
VT Viola Tamási
ZR Zsolt Rónai
MC Miklós Csala
ÉK Éva Kereszturi
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Rs1054411 promoter polymorphism of the SCD1 gene was genotyped using pre-designed TaqMan assay (C_34192814_10, Thermo Fisher Scientific, Waltham, MA, USA). qPCR assay was performed in 5 µL final volume containing approximately 4 ng genomic DNA, 1 × TaqPath™ ProAmp™ Master Mix, and 1 × TaqMan® SNP Genotyping Assay using QuantStudio 12 K Flex Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA). Thermocycle was started by activating the hot start DNA polymerase and denaturing genomic DNA at 95 °C for 10 min. This was followed by 40 cycles of denaturation at 95 °C for 15 s, and combined annealing and extension at 60 °C for 1 min. Real-time detection was carried out during the latter step to verify the results of the subsequent post-PCR plate reads and automatic genotype calls.

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