RNA preparation, labeling, hybridization on a HT12 array (Illumina), and scanning were performed according to Illumina instructions by Genomics Lab, Wellcome Trust Centre for Human Genetics (Univeristy of Oxford).

Protocols were developed to measure human immune responses to melanoma antigens to be measured using ELISpot and tetramers. The antigens in question were also inserted into adenovirus, lentivirus and vaccinia vectors for re-stimulation studies.

These procedures have led to an improvement in processing of small RNA samples. Studies of 3’ IVT microarrays expression profiling are obtainable starting from 2 ng of total RNA.

Array construction: The yeast oligonucleotide array was constructed using the S. cerevisiae Genome Oligo Set ™ (Operon Technologies, CA, USA) composed of 6240 optimized oligonucleotides (70mers) each representing one yeast gene. Probe preparation and hybridization: We used the indirect labeling method. Briefly, the reactive amine derivative of dUTP, 5-(3- aminoallyl)-2?-deoxyuridine 5?-triphosphate (Sigma) was incorporated into cDNA using the Superscript II reverse transcriptase (Invitrogen) and oligo dT (Invitrogen) and random examers (Roche). After synthesi...