Table 1. Effect of spacer length in the DNA bridge on the ligation efficiency.a .
Yield (%) | ||||||
2h | 6h | 20h | ||||
BRIDGE OLIGO | LIGATION | ADENYLATE | LIGATION | ADENYLATE | LIGATION | ADENYLATE |
A0G | 0.4 | 12 | 5.8 | 17 | 19 | 9.9 |
A1G | 15 | 36 | 69 | 12 | 87 | 3.6 |
A2G | 31 | 37 | 84 | 7.3 | 91 | 2.9 |
A3G | 11 | 14 | 39 | 7.6 | 63 | 6.8 |
A4G | 2.8 | 13 | 20 | 11 | 48 | 4.6 |
A6G | 0.7 | 6.7 | 1.4 | 8.5 | 9.1 | 10 |
A8G | 0.0 | 6.0 | 0.0 | 6.6 | 0.8 | 6.8 |
A10G | 0.0 | 5.9 | 0.0 | 6.9 | 0.1 | 11 |
T1G | 9.9 | 66 | 51 | 41 | 87 | 6.0 |
T2G | 86 | 8.5 | 93 | 3.7 | 94 | 3.2 |
T3G | 31 | 32 | 84 | 7.0 | 92 | 2.6 |
T4G | 20 | 48 | 79 | 14 | 92 | 2.7 |
T6G | 10 | 44 | 59 | 24 | 88 | 3.6 |
T8G | 1.6 | 21 | 8.1 | 37 | 43 | 20 |
T10G | 0.0 | 10 | 0.6 | 19 | 6.0 | 30 |
HEG-G | 11 | 37 | 42 | 35 | 69 | 29 |
Reactions using dCHr8 were performed at 20°C using 3.3 µM [32P]-labeled dCHr8, 4.1 µM of capture oligo and 4.1 µM of the indicated bridging template. All bridging oligos contained dG in the pairing position across from the tethered dC.