Abstract

We developed a novel microfluidic method to integrate the entire workflow of single cell genetic analysis, including single cell capture, zero-loss nucleic

acid extraction and purification from low copy cells (1-105 cfu) in a microwell

and furthermore perform single cell real-time PCR in the same microwell. All these steps are carried out in a single well, avoiding sample loss during sample transfer. We further applied this “all-in-one-well” sample prep integrated PCR to an array of

nanoliter wells for single cells dPCR analysis. This sample prep integrated genetic testing method also has a potential to carry out dPCR using a raw sample with zero loss of nucleic acid when removing PCR inhibitors. It also has benefits of reducing the total raw sample volume to 1ul or less, and selectively removing DNA or RNA to enable high quality gene expression analysis.