C 904973 10 rs7412

DNA from buffy coat was isolated using the FlexiGene DNA AGF3000 kit (Qiagen, Valencia, CA, USA) on an AutoGenFlex 3000 workstation (Autogen, Holliston, MA. USA) and genotyping was carried out in the Genetics Unit-Parque Cientifico de Madrid (Madrid. Spain). Briefly, samples were spotted onto 384 plates using a Beckman BioMek 2000 automated liquid handler (Beckman High Wycombe, UK) and diluted in a mix consisting in TaqMan Genotyping MasterMix (Applied Biosystems, Foster City, California)) and a mixture of pre-made TaqMan SNP genotyping assays; C_3084793_20 (rs429358) and C_904973_10 (rs7412) (Applied Biosystems). qPCR reactions were made in a HT7900 Fast Real-Time PCR System (Applied Biosystems). SDS 2.4 software (Applied Biosystems) was used for genotype calling.

DNA isolation from whole blood was performed by using the FlexiGene DNA AGF3000 kit (Qiagen, Valencia, CA, USA) on an AutoGenFlex 3000 workstation (Autogen, Holliston, MA, USA), and genotyping was carried out in the Genetics Unit-Parque Científico de Madrid (Madrid, Spain). DNA samples were spotted onto 384 plates using a Beckman BioMek 2000 automated liquid handler (Beckman High Wycombe, UK) and were diluted in a mixture consisting of TaqMan Genotyping MasterMix (Applied Biosystems, Foster City, California) and a mixture of premade TaqMan SNP genotyping assays: C_3084793_20 (rs429358) and C_904973_10 (rs7412) (Applied Biosystems). qPCR assays were performed in an HT7900 Fast Real-Time PCR System (Applied Biosystems), and SDS 2.4 software (Applied Biosystems) was used for genotype calling. Samples with known genotypes as well as negative amplification blanks were included within each run to serve as positive/negative controls and assist in genotyping. This methodology has been used and described previously^{29 (link)}.