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Whole genome association analysis will be performed on the 1 million SNP (single nucleotide polymorphism) arrays, and additional association analysis will be performed on additional phenotypes.
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A leave-one-out cross validation and external validation was performed on additional cell lines and adenocarcinomas.
In addition, gene-expression analyses (Real time PCR) were performed on additional samples.
In the Right Panels qPCR analysis was performed on additional targets in adult keratinocytes and two targets -TIG1 and Hes1- were not validated in the core promoter region considered.
Insulin resistance assays were performed on additional cohorts of 6 mice per genotype, for young (2 to 5 month) and old (18 month) males and young (2 to 5 month) females.
Immunohistochemistry for GOLPH3 was performed on additional 218 RCC tissues.
Since these established skeletal phenotypes are unambiguous, no bone analyses were performed on additional cohorts of mice.
Light microscopy and specialized image analysis to visualize crimping was performed on additional control and surgical specimens.
Q-PCR analysis was performed on additional 26 matched ESCC and adjacent non-neoplastic tissues for selected lncRNAs.
When significant association was observed in animal set 1, containing Rambouillets from the original GWAS (White et al. 2012), genotyping was performed on additional animal sets.
In order to gain further insight into temporal variation in isoform expression, the qRT-PCR was performed on additional embryonic and placental stages, ranging from E6 to E11.
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