Figure 3.

Influence of supplemental PEG-8000 and incubation temperature on the formation of adaptor–target heterodimer versus target homodimer. The standard ligation reaction in the microSAGE protocol is performed in the presence of 5% PEG-8000 (w/v) at 16°C for 2 h using a final adaptor concentration of 0.08 μM in a final volume of 20 μl. Ligation reactions shown were performed in a final volume of 10 μl as described in Materials and Methods using a adaptor concentration of 1 μM final in the presence or absence of PEG-8000 supplemented to a final concentration of 15% (w/v). The reactions were carried out for 2 h under the conditions indicated.