Below is a detailed description of how specimens are processed and stored.
PBMCs (EDTA unless otherwise noted in database)
All PBMCs are processed 1:1 with Sigma-Aldrich Ficoll Histopaque 1077 and washed with PBS before being counted and stored in freezing medium (50% RPMI, 40% FBS, 10% DMSO). PBMCs are in –80 freezer overnight then transferred and stored in LN2 for long term storage.
Plasma (100% – EDTA unless otherwise noted in database)
Whole EDTA blood tubes spun down for plasma at 1300 RPM. Then plasma siphoned off and spun down at 3000 RPM, which is then divided into 1.8mL aliquots and stored in –80.
Four milliliter of blood are removed and saved into 4mL EDTA tubes., which are then frozen at –20 overnight then transferred to –80 for long term storage. These samples may be thawed later for DNA extraction.
Study volunteer blood are drawn directly into PAXgene RNA tubes and frozen at –20 overnight then transferred to –80 for long term storage.
RNALater/Snap Frozen Skin/Tumor Biopsy
Skin/tumor biopsies are obtained and trisected for RNALater storage (after 2008) which is in 4C overnight then transferred to –80 for long term storage. All skin/tumor biopsies prior to RNALater are snap frozen in liquid nitrogen and stored in LN2.
OCT Skin/Tumor Biopsy
Skin/tumor biopsies are obtained and trisected for OCT storage. Samples are frozen in OCT with dry ice then trimmed of excess OCT to fit into 1.8mL Nunc Tube, which is then stored in –80 for long term storage.
Formalin Fixed Skin/Tumor Biopsy
Skin/tumor biopsies are obtained and trisected for formalin fixed storage. Samples are stored in 4C for long term storage.
Please note: The numbers below are ballpark figures.
Please note: Only ONE skin/tumor biopsy obtained per patient (unless otherwise noted), and the one sample is then trisected for RNALater, OCT and formalin storage. If you need further clarification or more information, please contact Helen Kuo (firstname.lastname@example.org).