Luminex 200 User Manual with LDS 1.7 Software - Manual (Page 126)

Luminex 200 User Manual for LDS Version 1.7
x
MAP Technology
9 - 8 PN 89-00002-00-150 Rev. A
• SDS:
Sodium Dodecyl Sulfate (0.1% w/v)
Preparation
1. Allow all reagents to warm to room temperature.
2. Resuspend the amino-substituted oligonucleotide to 1.0 mM in
sterile, deionized water.
Procedure
1. Centrifuge the xMAP microsphere stock for 1 minute at a
minimum of 8,000 x g.
2. Disperse the pellet with sonication, and vortex the container for
20 seconds.
3. Dispense 5.0
× 10 6
of the xMAP microspheres from the stock
tube into a 1.5 ml microcentrifuge tube (See Technical note 2).
4. Microcentrifuge the xMAP microspheres at 8,000
× g or higher
for 1 minute.
5. Remove the supernatant, being careful not to disturb the pellet.
6. Add 50 µl of 0.1 M MES (pH 4.5). Vortex and sonicate.
7. Add 0.2 nmole of amino-substituted oligonucleotide (i.e., 2 µl of
a 1:10 dilution of the 1 mM solution). Vortex briefly (See
Technical note 3).
8. Immediately before use, prepare a 10 mg/mL solution of EDC
powder (See Technical note 4). Vortex until dissolved.
9. Add 2.5 µL of the fresh EDC solution to the xMAP
microspheres. Vortex immediately.
10. Incubate for 30 minutes at room temperature in the dark.
11. Repeat steps 8-10 with fresh EDC (for a total of two EDC
additions).
12. Add 1.0 ml of Tween 20 (0.02% v/v). Vortex.
13. Centrifuge the xMAP microspheres at 8,000
× g or higher for 1
minute.
14. Remove the supernatant, being careful not to disturb the pellet.
15. Add 1.0 ml of SDS (0.1% w/v). Vortex.