A Single PDMS Based Chip for E. Coli Bacteria Lysis, DNA Extraction and Amplification

An integrated microfluidic device with on-chip performance of cell lysis, DNA extraction and PCR amplification is achieved to realize the total analysis from cell lysis to DNA amplification on a single polydimethylsiloxane (PDMS) chip. Using multilayered PDMS patterning and solid phase extraction (SPE) techniques, DNA is extracted by magnetic beads (MBs) from the cell lysate prepared by a two-step thermal regime. After DNA extraction, instead of traditional DNA amplification requiring washing MBs out, a non-elution on-chip PCR is performed with DNA captured by MBs serving directly as the template. The procedure from DNA extraction to amplification is simplified and can be realized on one chip without washing MBs out. The feasibility of realizing whole process on a single chip is verified by the gel electrophoresis result. This study shows an approach towards the integrated microsystem which is capable of comprehensive genetic analysis.