Kulinski, M. Dominika and Mahalanabis, Madhumita and Gillers, Sara and Zhang, Jane Y. and Singh, Satish and Klapperich, Catherine M. (2009) Sample preparation module for bacterial lysis and isolation of DNA from human urine. Biomedical Microdevices, 11 (3). pp. 671-678.
Silica impregnated polymer monolithic columns may provide a simple method for lysing and extracting DNA from bacteria inside of microfluidic chips. Here we use Escherichia coli as a test organism for a point of care thermoplastic microfluidic module designed to take in a urine sample, mix it with lysis buffer, and perform a hybrid chemical/mechanical lysis and solid phase extraction of nucleic acids from the sample. To demonstrate proof-of-concept, we doped human hematuric urine samples with E. coli at concentrations ranging from 101–105 colony-forming units/mL (CFU/mL) to simulate patient samples. We then performed on-chip lysis and DNA extraction. The bacterial DNA was amplified using real-time PCR demonstrating lysis and isolation down to 101 CFU/mL. Results were comparable to a commercial kit at higher concentrations and performed better at recovering DNA at lower concentrations.
|Subjects:||Biomedical Science > Nanobiotechnology|
Material Science > Nanochemistry
|Deposited On:||26 Aug 2010 08:00|
|Last Modified:||26 Aug 2010 08:00|
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