Implementation of Emulsion PCR for Amplification of Click-Modified DNA During SELEX.

Abstract

Biased amplification of enriched DNA libraries is a limitation in the SELEX process and reduces the chances for successful enrichment of target-binding sequences. Implementation of emulsion PCR into click-SELEX protocols for targeting proteins or cells prevents the formation of by-products and increases the probability of successful enrichment of binding sequences. Through compartmentalization even poorly amplifiable sequences can be enriched, and by-products formed by product-product or product-primer hybridization are reduced to a minimum. In this chapter, we describe a protocol for emulsion PCR and subsequent DNA recovery for implementation into click-SELEX protocols using click-modified DNA. Our emulsion PCR protocol is easily integrated into existing SELEX protocols, requires no special laboratory equipment, and can be performed with easily commercially available reagents.