Association between the rs1544410 polymorphism in the vitamin D receptor (VDR) gene and insulin secretion after gestational diabetes mellitus v1

Genotyping using TaqMan allelic discrimination assay Genotyping was done by adding 2 μl (total 10 ng) into MicroAmp™ Optical 384-well plate (Applied Biosystems, Sweden) and dried overnight. On the following day, a mixture of 2.5 µl of TaqMan® Genotyping Master Mix (Applied Biosciences, Sweden), 0.0625 µl of TaqMan® SNP Genotyping Assay (Applied Biosystems, Sweden) and 2.4 µl of sterile H2O was added to the wells using a multichannel pipette. The plates were covered with MicroAmp™ optical Adhesive Film (Applied Biosystems, Sweden), vortexed and spun down shortly. Plates were then placed in a thermal cycler (Applied Biosystems, Sweden) with following PCR conditions: 50°C for 2 min, 95°C for 10 min and 40 cycles of (95°C for 15 sec and 60°C for 1 min). The plates were read using QuantStudio™ 7 Flex Real-Time PCR System (Applied Biosystems, Sweden).TaqMan® SNP Genotyping Assay were designed using Custom Assay TaqMan® Design Tool (Applied Biosystems, Sweden). More details about the studied TaqMan® SNP are listed in the Table below. Chr.12: 57764205 on GRCh38