title: Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR.
abstract: Agarose emulsion droplet microfluidic technology for single copy emulsion PCR (ePCR) is a suitable technique for the detection of single copy DNA molecules. It improves the traditional ePCR by employing agarose with low melting and low gelling temperatures, which is coupled with PCR forward primers using Schiff-base reaction. Highly uniform monodisperse nanoliter agarose droplets each containing PCR reagents and single copy template are produced with a microfabricated emulsion generator. Following PCR, the cooled droplets transform to microbeads carrying amplicons to maintain the monocolonity of each droplet, which can be further analyzed. This method allows high-throughput generation of uniform droplets and enables high PCR efficiency, making it a promising platform for many single copy genetic studies.
authors: Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, People's Republic of China.
authors: Xuefei
authors: X
authors: Leng
authors: Chaoyong James
authors: CJ
authors: Yang
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004655:Emulsions; D005898:Glass / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D012545:Schiff Bases / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D004655:Emulsions; D012545:Schiff Bases; D012685:Sepharose