Fabrication of the thermoplastic microfluidic channels.
In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using polymer/nanoparticle composite microscale lysis and solid phase extraction columns. The recovered samples are concentrated, small volume samples that are PCRable, without any additional cleanup. Here, we demonstrate how to fabricate thermoplastic microfluidic chips using hot embossing and heat sealing. Then, we demonstrate how to use in situ light directed surface grafting and polymerization through the sealed chip to form the composite solid phase columns. We demonstrate grafting and polymerization of a carbon nanotube/polymer composite column for bacterial cell lysis. We then show the lysis process followed by solid phase extraction of nucleic acids from the sample on chip using a silica/polymer composite column. The attached protocols contain detailed instructions on how to make both lysis and solid phase extraction columns.
Bhattacharyya
Arpita
A
Department of Biomedical Engineering, Boston University, USA. abhatta@bu.edu
Kulinski
Dominika
D
Klapperich
Catherine
C
D001419:Bacteria / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry; D006361:Heating; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D055616:Microtechnology / Q000295:instrumentation / Q000379:methods; D037742:Nanotubes, Carbon / Q000737:chemistry; D010969:Plastics / Q000737:chemistry*; D013499:Surface Properties
D004269:DNA, Bacterial; D037742:Nanotubes, Carbon; D010969:Plastics