Luminex 100 User Manual Version 1.7 - Manual (Page 118)

Luminex 100 User Manual Version 1.7 x MAP Technology
9 - 8 PN 89-00002-00-063 Rev. A
One-Step
Carbodiimide
Coupling of
Oligonucleotides to
xMAP Carboxylated
Microspheres
Introduction
These protocols are a general starting point for developing assays.
All assays should be optimized for your reagents in your specific
application. Methods that give you the best results start with these
guidelines and are modified 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,
Pierce Chemicals
• 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.