Application Notes
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Example Protocol to Conjugate Malachine Green-NHS Ester to Oligonucleotides. (The same methods apply for conjugation of Leuco-Malachite Green NHS Ester to amines).The succinimide ester of Malachite Green (21 mg, 30.11 micromol) is dissolved in 1.0 mL methyl sulfoxide. The amino-oligo is then dissolved in 800 microL of 0.2 M sodium carbonate buffer (pH 9.5). The ester solution (400 microL) is added to the amino-oligo solution. The mixture is then left for 16 hour at 4 degrees C, after which, it is chromatographed on a Sephadex G-25 column using deionized water/carbonate buffer (50/50) as eluent. The fraction with deep blue coloration is dialyzed against water to remove excess salt and unreacted reagents, and then freeze-dried. The final product is stored in the refrigerator until use.
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