Luminex 100 IS Version 2.1 - Manual (Page 77)

PN 89-00002-00-062 Rev. A 71
x MAP Technology
One-Step Carbodiimide Coupling of Oligonucleotides to xMAP
Carboxylated 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
• Sonicator bath
• Micropipetters: (1 µL - 1000 µL)
• Microcentrifuge
• Timer
• Analytical balance
Materials
• xMAP carboxylated microspheres—LIMIT EXPOSURE TO
LIGHT!
• Amino-substituted oligonucleotide (See Technical note 1)
• Microcentrifuge tubes: 1.5 mL, polypropylene
• Pipetter tips: 1 µL - 1000 µL
• EDC:
1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride
• MES:
0.1 M 2- (N-morpholino) ethane sulfonic acid pH 4.5, filter and
store at 4°C.
• Tween 20 (0.02% v/v)
• 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. Disperse the pellet with sonication, and vortex the container for
20 seconds.
2. Dispense 5.0 × 10 6
of the xMAP microspheres from the stock
tube into a 1.5 mL microcentrifuge tube (See Technical note 2).
3. Microcentrifuge the microspheres at ≥ 8,000 × g for 1 minute.