Procedure for Stabilization
of Peripheral Blood or Bone Marrow
This procedure is for the stabilization of peripheral blood or bone marrow samples for use on the Signature OncoChip™. The amount of sample necessary is 150µl of peripheral blood or bone marrow (or the amount equivalent to 2-6 million cells if cell count is available). Please follow the procedure and use specimen kit contents provided by Signature Genomics.
Stabilization of samples using the following procedure should be done as soon as possible (within 24–48 hours), as the white blood cells can rapidly degrade, rendering poor results. Stabilizing the sample in cell lysis solution is a short procedure (about 15 minutes with incubation time) and is crucial for getting good quality, high molecular weight DNA for use on the microarray. Once the sample has been stabilized, it can be stored at room temperature and shipped to Signature Genomics for extraction and processing.
Cell Lysis Procedure
- Fill a microcentrifuge tube with 900µl of RBC Lysis Solution.
- Using a 1 ml syringe, draw 150µl (or the amount equivalent to 2–6 million cells if cell count is available) of whole blood or bone marrow into the syringe, and add to the tube containing RBC Lysis Solution.
- Invert multiple times and tap ends of two tubes together for uniform mixing. After all tubes have been inverted, incubate 10 minutes at room temperature, inverting again at least once during the incubation.
- Centrifuge at 13,000–16,000 x rpm for 30 seconds to pellet the white blood cells. Remove supernatant with a pipette and a filtered pipette tip, leaving behind the visible white pellet and about 10–20µl of liquid. Discard the supernatant into a beaker containing a 10% bleach solution.
- Vortex tubes vigorously to resuspend cells (greatly facilitates cell lysis step).
- Resuspend cells by adding 300µl cell lysis solution to each tube and vortex vigorously to lyse cells.
- Ship samples at room temperature to Signature Genomics.