Gel-Free Electrophoresis of DNA and Proteins on Chips Featuring a 70 nm Capillary–Well Motif

We present an integrated glass capillary system on silicon for size-based sieving of distinct mixtures of proteins, short DNA, and long DNA fragments into sharp peaks. The minimum resolvable size difference achieved is noted as 3.45 kDa for 45-52.8 kDa proteins, 20 bp for 200-300 bp DNA strands, and 182 bp for 5.6-5.8 kbp DNA chains. This high-resolution sieving arises from vastly steep entropic barriers created at the onsets of extremely restrictive (resistive) capillary segments and their pivotal role in shifting the equilibrium entropic sieving to intense fields (>1000 V/cm). DNA fragments of various sizes are shown fully resolved in less than 7 min at a steady voltage of 2000 V being directly applied across the length of a 2 cm long sieve featuring thousands of entropic barriers. The utility of higher field strengths and longer sieves is also demonstrated without triggering dielectric breakdown by time-division multiplexing up to 2000 V across the 1 cm long sieve segments. The self-enclosed 70 nm diameter capillaries were fabricated using coarse (>1 μm) photolithography and standard semiconductor manufacturing techniques.