Electrokinetically Integrated Isolation and Amplification of Protein-binding Nucleic Acids on a Microchip

This paper presents a microchip that integrates the isolation and amplification of target-binding DNA strands in a randomized DNA mixture. Target-binding strands are isolated in the chip via binding with human immunoglobulin E (IgE) immobilized on microbeads, electrophoretically transported through a gel-filled microchannel, captured onto microbead-tethered reverse primers, and amplified using polymerase chain reaction (PCR). Integration of isolation and amplification is achieved using electrophoretic transport through the gel-filled microchannel, which prevents contamination of buffers or reactants while transferring the desired target-binding DNA. Experimental results show that our microchip can isolate and amplify target-binding DNA strands with increased binding affinity to IgE protein.