Cell Lysis and High Molecular Weight DNA Extraction from Eukaryotic and Prokaryotic Cells Powered by AFA-EngergeticsTM
This application note describes a novel automatable approach to cell lysis and extraction of high molecular weight (HMW) DNA (>30,000 base pairs) from microbial samples, which can be readily sequenced with long-read sequencing technologies such as PacBio or Oxford Nanopore. Lysing the cells and subsequent extraction of HMW DNA from microbial samples is achieved on a Covaris LE220-plus Focused-ultrasonicator using a non-contact method mediated by AFA. Rapid lysis is achieved in less than 2 minutes for 96 individual bacteria samples and 10 to 60 minutes for 96 individual yeast samples when using a PCR-compatible 96 oneTUBE-10 AFA Plate. Both electropherograms and Qubit measurements reveal that high molecular weight DNA can be efficiently extracted using AFA-mediated cell lysis.