Figure 1:

Isolation of ultrapure platelet mRNA. A) Leukocyte depletion using CD45-coated magnetic beads resulted in only minimal loss of platelets from PRP in comparison to leukocyte depletion using PALL filtration. B) Increasing volumes of CD42b-coated magnetic beads enabled positive selection of ~75% of platelets from 1.5 ml of PRP. C) To test for purity of platelet sample preparations, platelet mRNA was reverse transcribed and amplified for detection of cell-specific markers, GPIBB (platelets), CD45 (PTPRC) (leukocytes) and GYPA (erythrocytes), and expressed relative to the endogenous reference gene GAPDH. D) Melt curve analysis for platelet (GPIBB), leukocyte (CD45) and erythrocyte (GYPA) in platelet sample preparations (top row) used for channelome screening, and in leukocyte (middle row) and erythrocyte (bottom row) sample preparations for comparison.