The Facility for Geroscience Analysis has standard preparation steps for its samples. Below are the preparation steps and information for urine, serum, and platelet poor plasma. Also, each preparation type has a downloadable pdf version of the figure and steps. If you have any questions, please contact us.
Step 1: After collection, keep urine refrigerated/on ice until processed.
Step 2: Transfer urine to a clean tube.
Step 3: Centrifuge the urine tube at 1400 x g for 10 minutes at 4°C
Step 4a: Transfer urine supernatant to a new clean tube
Step 4b: Cell pellet can be saved for future use. It contains cells and other elements.
Step 5: Aliquot urine supernatant into 1.5ml aliquots until the sample is exhausted.
Step 6: Store Aliquots at -80 °C until used.
Serum can be prepared from whole blood when collected on a tube without anticoagulant, containing or not clot activator. The blood clot can also be collected and frozen for future assays as it is a good source of DNA without preservatives.
Step 1: After the blood draw, keep the blood tube at room temperature for 30 minutes to allow clot formation.
Step 2: Centrifuge the blood tube at 2000 x g for 10 minutes at 4°C
Step 3a: Transfer the serum supernatant to a new clean tube and mix by inverting five times
Step 3b: Optionally the blood clot can be transferred to a new tube and frozen for future use.
Step 4: Aliquot serum into 0.5ml aliquots until the sample is exhausted.
Step 5: Store Aliquots at -80 °C until used
Platelet Poor Plasma Preparation and Buffy Coat Preparation
Platelet-poor plasma and buffy coat can be prepared from whole blood with EDTA, Heparin, or Citrate/ACD as an additive. Buffy coat can be prepared for future use of live cells or not, depending on the assay to be run with those samples.
Platelet-Poor Plasma Preparation
Step 1: After the blood draw, keep/transport blood tube refrigerated/on ice until processed.
Step 2: Centrifuge at 2000 x g for 10 minutes at 4°C
Step 3a: Transfer plasma to a new tube
Step 3b: Collect buffy coat
Step 4: Centrifuge plasma tube at 2000 x g for 10 minutes at 4°C
Step 5: Transfer Plasma Supernatant to new tube Mix by inverting 5 times.
Step 6: Aliquot EDTA/Hep platelet-poor plasma into 0.5ml aliquots until the sample is exhausted.
Buffy Coat Preparation
Step 3b: Transfer Buffy Coat into a 1.5 ml cryovial (Note: If no live cells are needed, freeze the tube at this time)
Step 3c: Add an equal volume of Freezing media and aliquot into two cryovials.
Step 3d: Slow-freeze overnight at -80 oC
Step 3e: Store in liquid Nitrogen