Table 2.
Estimated Labeling Efficiencies at Different MPc/oligonucleotide Molar Concentration Ratios Using Different Dyes, Linkers, and Succinimidyl Estersa
| Linkerb | MPc | succinimidyl ester | MPc/oligonucleotide | labeling efficiency |
|---|---|---|---|---|
| C12H24 | 1 | 3 | 5:1 | <5% |
| C12H24 | 1 | 3 | 10:1 | 24 ± 6% |
| C12H24 | 1 | 3 | 18:1 | 48 ± 2% |
| C12H24 | 2 | 5 | 11:1 | 78 ± 5% |
| C12H24 | 2 | 5 | 21:1 | 56 ± 3% |
| C12H24 | 2 | 5 | 38:1 | 39 ± 2% |
| C6H12 | 2 | 4 | 1:1 | <5% |
| C6H12 | 2 | 4 | 4:1 | 28 ± 3% |
| C6H12 | 2 | 4 | 9:1 | 38 ± 6% |
| C12H24 | 2 | 4 | 1:1 | 33 ± 6% |
| C12H24 | 2 | 4 | 4:1 | 47 ± 3% |
| C12H24 | 2 | 4 | 11:1 | 53 ± 4% |
a
Conjugation reactions were performed in borate buffer at pH 8.5 and using an oligonucleotide concentration of 20 nM.
b
The aliphatic linker used to attach the amino group to the 5′ end of the oligo-nucleotide.