Microfluidic DNA fragmentation for on-chip genomic analysis

作者：Shui Lingling^*; Bomer Johan G; Jin Mingliang; Carlen Edwin T; van den Berg Albert

来源：Nanotechnology, 2011, 22(49): 494013.

DOI：10.1088/0957-4484/22/49/494013

• Deoxyribonucleic acids
• DNA fragmentation
• Fragment lengths
• Genomic analysis
• High-throughput
• Hydraulic pressure
• Hydrodynamic pressure
• Microfluidic channel
• Micromachined
• On chips
• Recirculations
• Velocity gradients

We report a high-throughput clog-free microfluidic deoxyribonucleic acid (DNA) fragmentation chip that is based on hydrodynamic shearing. Salmon sperm DNA has been reproducibly fragmented down to similar to 5k bp fragment lengths by applying low hydraulic pressures (<= 1 bar) across micromachined constrictions positioned in larger microfluidic channels that create point-sink flow with large velocity gradients near the constriction entrance. Long constrictions (100 mu m) produce shorter fragment lengths compared to shorter constrictions (10 mu m), while increasing the hydrodynamic pressure requirement. Sample recirculation (10x) in short constrictions reduces the mean fragment length and fragment length variation, and improves yield compared to single-pass experiments without increasing the hydrodynamic pressure.

• 出版日期2011-12-9