Standard Protocol for Collection and Storage of Plasm
The method below is based on the method BD has developed, with a few small
changes in the centrifugation step, etc.
1. Please refer to the P100 package insert (attached) for venipuncture techniques.
2. The BD P100 tubes are to be stored at 4C, and should be prechilled at 4C (not frozen) until use.
3. A full 8.5 mL of
blood should be collected. This will produce about 2.5-3 mL of plasma.
4. Immediately after collection, invert the tube 8-10
times to mix the protease inhibitors and anticoagulent with the blood sample.
5. Place tube in wet ice before centrifuging.
(Centrifugation should be done within 30 minutes of collection).
6. Centrifuge at 2000-3000 RCF at 4C for 15 min. (See BD P100
package insert for converting rpm to RCF). Do not exceed 3000 g, or 10,000 RCF.
7. Within 30 minutes of centrifugation, transfer
the plasma in 1-mL aliquots to pre-labeled Fisherbrand 4-mL self-standing cryovials (Fisher Scientific # 0566966).
8. Place
aliquots immediately on dry ice.
9. Freeze aliquots at –80C until used. (Avoid freeze-thaw cycles).
To remove microplatelets, the plasma should be transferred to a different centrifuge tube, and centrifuged at 12,000 g at 4C for 15 min.
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Standard protocol for collection and storage of plasm
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