Why is there an extra Buffer RPE wash in the post-IVT cleanup?
In the post-IVT cleanup, the Qiagen RNeasy kit is used to purify RNA from the IVT reaction mixture. The protocol followed is similar to the RNA reaction cleanup protocol provided by the kit handbook, except for an extra Buffer RPE wash. We have found this wash to be necessary if the RNA is to be used for subsequent microarray applications involving fluorescence, since it is necessary to remove any trace GITC salts that may otherwise remain and produce undesirable background fluorescence on the microarrays.
