Sample Preparation and Submission
Requirements Before Submitting RNA Samples:
RNA from mammalian cells: Affymetrix recommends using Qiagen’s RNeasy Total RNA isolation kit.
RNA from mammalian tissue: Affymetrix recommends isolating total RNA with a commercial reagent such as TRIzol.
All samples MUST be cleaned up using the Qiagen RNeasy Mini Kit or an equivalent column-based cleanup before turning them into this facility. RNA quality should be checked before turning the RNA into us. This includes looking at the 260/280 ratio and running a Tapestation or Bioanalyzer analysis. A quality check is required before proceeding with the microarray service. This facility can provide a TapeStation QC analysis if needed.
Requirements Before Submitting DNA Samples for Axiom Genotyping Plates
A minimum of 200 ng of DNA at a concentration of 10 ng/µl µl is required for each sample that is turned in. Starting DNA must be double-stranded for the purpose of accurate concentration determination. DNA must be of high purity. DNA should be free of DNA polymerase inhibitors. Examples of inhibitors include high concentrations of heme (from blood) and high concentrations of chelating agents (i.e., EDTA). The gDNA extraction/ purification method should render DNA that is generally salt-free because high concentrations of particular salts can also inhibit enzyme reactions. DNA purity is indicated by OD260/OD280 and OD260/ OD230 ratios. The OD260/OD280 ratio should be between 1.8 and 2.0 and the OD260/OD230 ratio should be greater than 1.5.DNA must not be degraded. The approximate average size of gDNA may be assessed on a 1% agarose gel using an appropriate size standard control. Approximately 90% of the DNA must be greater than 10 Kb in size.