Manipulation and extraction of genomic DNA from cell lysate by functionalized magnetic particles for lab on a chip applications.
A novel approach for extracting living cells' genomic DNA materials utilizing functionalized magnetic particles (MPs) is reported in this investigation. This strategy is amenable to handle bio-samples in a miniaturized environment and it offers a possibility to separate and purify DNA from other cell lysate mixtures "on-chip", which is known to be a bottle-neck step in an integrated micro-total-analysis-system (muTAS). "Species-specific" genomic DNA of interest is captured by the MPs based on the hybridization interaction between the biotinylated probes modified MPs and a complementary region of the targeted genome. The genome DNA anchored on the particles can be separated from the rest of cellular mixtures by a simple buffer washing upon the exertion of external magnetic force. Surface modifications of MPs and hybridization conditions affecting the genome capturing efficiency are investigated. Extraction of genomic DNA from E. coli is demonstrated in a silicon/glass-based micro-reactor patterned with a platinum heater and sensors. On-chip extraction and manipulation of genomic DNAs illustrated in this study is a step forward toward a total integrated bioanalytical microsystem for crude cells/sample analysis.
Yeung
Siu Wai
SW
Department of Chemical Engineering, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Hsing
I-Ming
IM
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics*; D016680:Genome, Bacterial; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods; D008280:Magnetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D010316:Particle Size
D004269:DNA, Bacterial