Luminex 200 User Manual with LDS 1.7 Software - Manual (Page 127)

x
MAP Technology Protocols
PN 89-00002-00-150 Rev. A 9 - 9
16. Centrifuge the xMAP microspheres at 8,000
× g or higher for 1
minute.
17. Remove the supernatant, being careful not to disturb the pellet.
18. Resuspend the xMAP microspheres in 100 µl of TE (10mM Tris,
1mM EDTA, pH 8.0).
19. Enumerate the xMAP microsphere preparation.
20. Store the preparation at 2
o
C - 8°C. Protect it from light.
Technical notes
1. Oligonucleotides must be synthesized with a 5’ or 3’
amino
group. No Tris or Azide or other amine-containing buffers
should be present during the coupling procedure. Preferably, the
oligonucleotide should be resuspended in water following
synthesis. If oligonucleotides were previously solubilized in an
amine-containing buffer, then precipitation and resuspension into
water is required.
2. This procedure can be scaled up or down.
3. The optimal coupling concentration for a given oligonucleotide
is determined by coupling at various concentrations within the
recommended range of 1 to 50 µM.
4. Minimize the exposure of EDC to air; secure closures on stock
and dry aliquot containers, store desiccated at -20°C. Use
aliquots immediately and discard containers after use. Make a
fresh 10 mg/ml EDC solution before each addition.
Binding Biotin-
conjugated
Molecules to
LumAvidin-Modified
xMAP Microspheres
Introduction
Use the protocols included here as a general starting point for
developing assays. All assays should be optimized for your reagents
in your specific application. You will discover the methods that give
you the best results by starting with these guidelines and modifying
them for your specific needs.
Equipment
• Vortex