Housekeeping Genes

Use this option to select variables corresponding to target sequences that are consistent in their expression levels and are either recognized by the CodeSet (marked as Housekeeping, for example), or according to the user’s prior knowledge. The choice of reference genes to include in the CodeSet is therefore a crucial part of experimental design and Nanostring recommends selecting at least three reference genes. However, you should keep in mind that the more reference genes you specify, the more accurate the normalization.

If Housekeeping Genes is specified as the Reference Genes to Use but no housekeeping genes are specified using this option, this process will use all of the available genes in the data set as reference genes. Note: This practice is consistent with the total.sum method in the NanoStringNorm R package.

To Specify the Housekeeping Gene Variables:

Specify an input data set.

Make sure that Housekeeping Genes is specified as the Reference Genes to Use.

All of the variables in the input data set are listed in the Available Variables field.

Left-click a single variable from the Available Variables field.

Click to add the highlighted variable to the Housekeeping Gene field.