Estimation of SELEX pool size by measurement of DNA renaturation rates

We report a general method of measuring the complexity of SELEX pools. In a nalogy to measurements of genome size by C(0)t analysis, the complexity of a SELEX pool is measured by determining the reannealing rate of its double- stranded PCR product. We applied this technique to study the selection dyna mics of a recently reported SELEX to neutrophil elastase. We found that the number of sequences decreased from 10(7) in round 6 to similar to 60 by ro und 15, the final round. The intermediate rounds are a mixture of a high ab undance/low complexity pool with a low abundance/high complexity pool. As t he SELEX progresses, the former pool expands at the expense of the latter. This technique should be useful for studying and optimizing SELEX dynamics, as well as for monitoring the progress of SELEX experiments.