KORA F4 samples were genotyped for the SNP rs1421085 with a concordance rate of >99.5% using the MassARRAY system with iPLEX chemistry (Sequenom, USA), as previously described by Holzapfel et al. (60 (link)). All subjects′ FTO rs1421085 genotype was validated by Sanger sequencing. The following primers were used: polymerase chain reaction (PCR) primers: 5′-accatcaaagaggctgttgt-3′ (rs1421085_PCRfwd) and 5′-gcacccattaactcgtcatt-3′ (rs1421085_PCRrev); sequencing primers: 5′-tgtctctaagcccaacaaac-3′ (rs1421085_Seqfwd) and 5′-attgagccatccatcaggtt-3′ (rs1421085_Seqrev).
The PCR was performed with around 50 ng of input genomic DNA in a thermocycler (Biometra, Jena, Germany) as follows: 12 min at 95°C, 50 cycles of 20 s at 95°C, 40 s at 56°C, and 90 s at 72°C, and finally 2 min at 72°C before cooling (8 (link)).
The PCR was performed with around 50 ng of input genomic DNA in a thermocycler (Biometra, Jena, Germany) as follows: 12 min at 95°C, 50 cycles of 20 s at 95°C, 40 s at 56°C, and 90 s at 72°C, and finally 2 min at 72°C before cooling (8 (link)).
Free full text: Click here
