DNA capture-probe based separation of double-stranded polymerase chain reaction amplification products in poly(dimethylsiloxane) microfluidic channels
Dmitriy Khodakov, Leigh Thredgold, Claire E Lenehan, Gunther G Andersson, Hilton Kobus, Amanda V Ellis
BIOMICROFLUIDICS | AMER INST PHYSICS | Published : 2012
Herein, we describe the development of a novel primer system that allows for the capture of double-stranded polymerase chain reaction (PCR) amplification products onto a microfluidic channel without any preliminary purification stages. We show that specially designed PCR primers consisting of the main primer sequence and an additional "tag sequence" linked through a poly(ethylene glycol) molecule can be used to generate ds-PCR amplification products tailed with ss-oligonucleotides of two forensically relevant genes (amelogenin and human c-fms (macrophage colony-stimulating factor) proto-oncogene for the CSF-1 receptor (CSF1PO). Furthermore, with a view to enriching and eluting the ds-PCR pro..View full abstract
The authors gratefully thank Professor Adrian Linacre for providing molecular biology facilities and the Queensland Government Smart State National and International Research Alliances Program for funding.