Abstract
The thermal expansion of gas and the air permeability of polydimethylsiloxane (PDMS) were previously thought to be the main causes of bubbles and water loss during polymerase chain reaction (PCR), resulting in a very complex chip design and operation. Here, by calculating and characterizing bubble formation, we discovered that water vapor is the main cause of bubbling. During PCR, heat increases the volume of the bubble by a factor of only ~0.2 in the absence of water vapor but by a factor of ~6.4 in the presence of water vapor. In addition, the phenomenon of "respiration" due to the repeated evaporation and condensation of water vapor accelerates the expansion of bubbles and the loss of water. A water seal above 109 kPa can effectively prevent bubbles in a bare PDMS chip with a simple structure, which is significant for the wide application of PDMS chips.