Bio-Rad Nonmagnetic Beads and Related Reagents - Manual (Page 25)

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20. Place the tube into a magnetic separator and allow separation to occur
for 30–60 sec.
21. With the tube still positioned in the magnetic separator, carefully remove
the supernatant without disturbing the bead pellet.
22. Remove the tube from the magnetic separator and resuspend the
coupled beads in 100 µl of TE, pH 8.0, by vortex and sonication for
approximately 20 sec.
23. Enumerate the coupled beads by hemacytometer:
a. Dilute the resuspended coupled beads 1:100 in dH
2
O.
b. Mix thoroughly by vortex.
c. Transfer 10 µl to the hemacytometer.
d. Count the beads within the 4 large corners of the hemacytometer
grid.
e. Beads/µl = (sum of beads in 4 large corners) x 2.5 x 100 (dilution
factor).
f. Note: maximum is 50,000 beads/µl.
24. Store coupled beads refrigerated at 2–8°C in the dark.