Labeled cDNA from each of the eight time points was hybridized to a microarray slide containing 8 separate copies of the E. coli genome (Agilent Microarray Design ID 020097) according to instructions provided by the manufacturer.
The whole volume of the purified probe was denatured at 95°C for 2 min and was applied to the cDNA microarray slide containing 8464 genes, which was provided by United Gene Holdings (Biostar Genechip Inc., Shanghai, China).
Samples were hybridized on a microarray slide containing almost 44,000 probes per array coding for ~ 14,000 gene transcripts indicating that for a subset of genes more than one probe was present.
To further survey the PCYT1A protein tissue expression profile, IHC analyses were performed on a tissue microarray slide containing 12 normal human tissue samples and 12 corresponding human tumor tissue samples.
The inter-observer variability of TOPK scores was assessed on one tissue microarray slide containing 456 cases by a second independent pathologist (MH) from an external institution and blinded to clinicopathological features.
Each Cy3- (or Cy5-) coupled PAM sample was then hybridized to its corresponding Cy5- (or Cy3-) coupled reference sample anchored on a pig microarray slide containing 20,400 oligonucleotides (US Pig Genome Coordination Program; http://www.animalgenome.org/pig/resources/array_request.html).html
