PCaP Collection, Processing and Storage Procedures
Blood: Approximately 42ml of blood was obtained from consenting participants, including three 8.5ml yellow top (ACD) tubes, one 10 ml red top tube, and one 6.5ml lavender top (EDTA) tube. Red and lavender top tubes were wrapped in foil and were transported on ice prior to initial processing. Serum was removed from the red top tube and aliquoted into ten cryovials. Lavender top samples were processed into plasma (six aliquots) and packed red blood cells (two aliquots). Yellow top tubes were transported at room temperature, and Louisiana samples were shipped overnight to UNC for processing and lymphocyte immortalization. Immortalized lymphocytes were divided into 6 aliquots and cryopreserved in liquid nitrogen. Plasma was removed from yellow-top tubes and DNA was purified from white blood cells. DNA, plasma, serum and packed red blood cells were stored long-term at -80°C.
Buccal rinse: For participants whose DNA was unavailable from blood, a retrospective collection of buccal rinse samples was conducted in NC and LA. Beginning on September 4, 2007, LA participants who, at the time of the study visit, could not complete the blood draw for DNA were given the option to complete a buccal rinse instead.
Urine: A 20ml urine sample was requested from participants. The study nurse immediately aliquoted half of the sample into a 15ml conical centrifuge tube containing 20 mg of crystalline ascorbic acid (as a preservative) and placed the remainder into a second conical tube without preservative. Samples were wrapped in foil and transported on ice prior to long-term storage at -20°C.
Adipose tissue: Subcutaneous fat samples were obtained from the abdominal area of consenting participants who were not allergic to local anesthetics. After the overlying skin was anesthetized with 2% lidocaine solution, a 15-gauge needle was inserted into the subcutaneous fat and suction was applied using a 15ml vacutainer tube. Aspirated tissue was trapped in the needle and Luer lock adapter, which were placed in separate cryovials, transported on ice, and stored at -80°C.
Toenail clippings: Participants were asked to collect toenail clippings from each toe of one foot prior to the study visit. Toenails were stored in a cryovial at ambient temperature.
Diagnostic and Radical Prostatectomy Tissue: See Tumor Block Retrieval and Tissue Microarray (TMA) Construction.
