title: Microfluidic chip for stacking, separation and extraction of multiple DNA fragments.
abstract: A disposable integrated microfluidic device was developed for rapid sample stacking, separation and extraction of multiple DNA fragments from a relatively large amount of sample. Isotachophoresis hyphenated gel electrophoresis (ITP-GE) was used to pre-concentrate and separate DNA fragments, followed by extraction of pure DNA fragments with electroelution on-chip. DNA fragments of 200bp, 500bp and 1kbp were successfully separated and collected in the extraction chamber within 25min. The extraction efficiency obtained from the chip was 49.9%, 52.1% and 53.7% for 200bp, 500bp and 1kbp DNA fragments, respectively. The extracted DNA fragments exhibited compatibility with downstream enzymatic reactions, for example PCR. The chip was also used to extract DNA fragments with specific size range from sheared genomic DNA and demonstrated similar performance to that using traditional gel cutting method. The whole assay can finish in 32min, 6 times faster than traditional method.
authors.affiliation: Singapore Institute of Manufacturing Technology, 71 Nanyang drive, 638075, Singapore. Electronic address: rgwu@SIMTech.a-star.edu.sg.
authors.forename: Ruige
authors.initials: R
authors.lastname: Wu
authors.affiliation: Singapore Institute of Manufacturing Technology, 71 Nanyang drive, 638075, Singapore.
authors.forename: Y P
authors.initials: YP
authors.lastname: Seah
authors.affiliation: Singapore Institute of Manufacturing Technology, 71 Nanyang drive, 638075, Singapore.
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Wang
mesh_terms: D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D057625:Isotachophoresis; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation*; D008967:Molecular Biology / Q000295:instrumentation* / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA
keywords: Extraction; Gel electrophoresis; Isotachophoresis; Microfluidic; Separation