What is the best method to use for extracting RNA from FACS?
Please consult with the microarray staff for successful RNA recovery from cells sorted using flow cytometry. Several key points are highlighted below: Preparing the flow cytometry is not a small task as it must be completely free of RNases from the sheath tank to the sorting nozzle. This decontamination procedure will take considerable time, so be prepared. Ensure the dip tubes, septa, flow cell, all tubing lines, and nozzles have been completely decontaminated with bleach, RNase ZAP, ethanol, autoclaving, or other qualifying technique prior to the sort. Rinse cytometer with DEPC water. Remember DEPC water DOES NOT inactivate RNases, and is only RNase-free water. The sheath fluid and tank must be RNase free. Use only sterile RNase free tubes on the cytometry that have never been open to contaminated air. As a control, retain some cells and extract the RNA to determine the condition of RNA prior to the sort. Also before and after trypsinization for adherent cells. Often the flow cytomet
