Luminex 100 IS Developer Workbench Guide Version 2.3 - Manual (Page 64)

Luminex 100 IS Developer Workbench Guide Version 2.3
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MAP Technology
58 PN 89-00002-00-084 Rev. B
Enumerate the Coupled
Microspheres
1. Dilute the resuspended coupled microspheres 1:100 in deionized
water.
2. Mix thoroughly using a vortex.
3. Transfer 10 µL to a hemacytometer.
4. Count the microspheres within the 4 large corners of the
hemacytometer grid.
5. Microspheres/µL = (Sum of microspheres in 4 large corners) x
2.5 x 100 (dilution factor).
6. Store the preparation at 2
o
C - 8°C. Protect it from light.
Technical Notes
1. Synthesize the Oligonucleotides with a 5’ or 3’
amino group and
spacer modification. No Tris or Azide or other amine-containing
buffers should be present during the coupling procedure.
Preferably, resuspend the oligonucleotide in water following
synthesis. If oligonucleotides were previously solubilized in an
amine-containing buffer, then precipitation and resuspension into
water. Refer to http://luminexcorp.custhelp.com for more
information.
2. You can scale this procedure up or down. Refer to http://
luminexcorp.custhelp.com for more information.
3. The optimal coupling concentration for a given oligonucleotide
is determined by coupling at various concentrations within the
recommended range of 1 to 100 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.
5. For best results, use microcentrifuge tubes from USA Scientific,
Inc., catalog number 1415-2500. To order, call 1 800 LAB-TIPS.