Total RNA for Gene Expression Profiling

  1. Total RNA of very high quality (28S:18S = 2:1, OD 260/280 > 1.8, OD 260/230 > 1.8) in DEPC-H2O is the best way to begin.
  2. Samples should be cleaned with Qiagen Rneasy columns, including Trizol-isolated RNA. See protocols. Samples for micro RNA expression should be isolated and purified with Qiagen miRNeasy Mini kit (cat. no. 217004).
  3. Dnase-treatment is highly recommended as RNeasy columns alone may not completely remove genomic DNA. See protocols.
  4. Total RNA isolated from whole blood needs to be globin-reduced. We recommend Ambion GLOBINclear kit (cat. no. AM1980, AM1981).
  5. Samples that need to be concentrated at our facility will be assessed an additional fee.
  6. Illumina expression samples should be batched to match the capacity of each microarray. For example, the Hu6 array will accommodate 6 samples.
platformassaymin. amountmin. conc. in H2O
affymetrixIVT express> 3 ug> 75 ng/ul
affymetrixexpression (3'-standard)> 6 ug> 0.5 ug/ul
affymetrix2-cycle expression> 2 ug> 50 ng/ul
affymetrixexon> 4 ug> 750 ng/ul
affymetrixgene array> 2 ug> 150 ng/ul
exiqonmiRNA expression> 200 ng> 50 ng/ul
illuminaexpression> 3 ug> 75 ng/ul
illuminaexpression hyb only> 1.5 ug cRNA> 150 ng/ul
illuminamicro rna expression> 2 ug> 150 ng/ul
illuminadasl> 2 ug> 150 ng/ul
nimblegenexpression> 10 ug> 1 ug/ul
nimblegenexpression hyb only> 13 ug> 1 ug/ul
spottedexpression> 2 ug> 0.5 ug/ul
spottedexpression hyb onlyplease inquire> 0.5 ug/ul