normalization data transformation protocol
Relative quantification of gene expression was determined using the comparative cycle threshold method (2(-ddCt)) as previously described Ref : Livak, K. J., and T. D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402-408. . ID_REF = VALUE = Expression of target genes was normalized in relation to expression of GAPDH.
array scanning protocol
nucleic acid extraction protocol
All colorectal tissue samples were maintained at -80ºC until RNA extraction and grinded under liquid nitrogen. Homogenization was achieved using QIAshredder homogenizers and total RNA was extracted using RNeasy Mini kit (both kits from Qiagen Inc; Valencia, CA, USA).