An injection molded microchip for nucleic acid purification from 25 microliter samples using isotachophoresis.
We present a novel microchip device for purification of nucleic acids from 25μL biological samples using isotachophoresis (ITP). The device design incorporates a custom capillary barrier structure to facilitate robust sample loading. The chip uses a 2mm channel width and 0.15mm depth to reduce processing time, mitigate Joule heating, and achieve high extraction efficiency. To reduce pH changes in the device due to electrolysis, we incorporated a buffering reservoir physically separated from the sample output reservoir. To reduce dispersion of the ITP-focused zone, we used optimized turn geometries. The chip was fabricated by injection molding PMMA and COC plastics through a commercial microfluidic foundry. The extraction efficiency of nucleic acids from the device was measured using fluorescent quantification, and an average recovery efficiency of 81% was achieved for nucleic acid masses between 250pg and 250ng. The devices were also used to purify DNA from whole blood, and the extracted DNA was amplified using qPCR to show the PCR compatibility of the purified sample.
Department of Chemical Engineering, Stanford University, USA.
L A
LA
Marshall
Department of Mechanical Engineering, Stanford University, USA.
A
A
Rogacs
Department of Mechanical Engineering, UC Santa Barbara, USA.
C D
CD
Meinhart
Department of Mechanical Engineering, Stanford University, USA. Electronic address: juan.santiago@stanford.edu.
J G
JG
Santiago
D004247:DNA / Q000097:blood; D006863:Hydrogen-Ion Concentration; D057625:Isotachophoresis; D044085:Microfluidics; D009696:Nucleic Acids / Q000097:blood / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction
D009696:Nucleic Acids; D004247:DNA
Device design; Isotachophoresis; Microfluidic; Nucleic acids; Sample preparation