Illumina gene expression protocols feature first- and second-strand reverse transcription steps, followed by a single in vitro transcription (IVT) amplification that incorporates biotin-labelled nucleotides. Subsequent steps include array hybridization, washing, blocking, and streptavadin-Cy3 staining. Fluorescence emission by Cy3 is quantitatively detected for downstream analysis.
Pricing: Please inquire
Pricing is highly dependent on experimental design - please allow us to help you find the most cost effective solution, take advantage of our complimentary experimental design consultation!
High quality total RNA is required for microarray experiments.
We require a 260/230 ratio of greater than 1.8 and a Bioanalyzer RIN of >7.5 in order to proceed with the gene expression array experiment.
Please note that if samples do not meet the required quality standards the project will be stopped and a fee of $10/sample (to cover the sample quality assessment) will be charged. The PMGC also offers several options for dealing with degraded RNA, however, this may require a change of platform and a new quotation to be issued. We will work with you to choose the best approach if you are unable to obtain RNA of sufficient quality.
For standard Illumina array processing, we require a minimum of 200 ng of total RNA at >40 ng/µl. This allows for processing of the sample as well as quality control tests. If you do not have sufficient RNA, please contact us to discuss other amplification strategies.
Partially degraded samples (such as from FFPE samples) can be labelled using the DASL Assay (cDNA-mediated annealing, selection, extension, and ligation assay). Please contact us for details of sample requirements for this assay.