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.
Extraction of genomic DNA (gDNA) from whole blood is the first step in multiple translational research and molecular diagnostics applications, such as next-generation sequencing (NGS), multiplex PCR, qPCR, and droplet digital PCR (ddPCR). For example, DNA from whole blood is frequently used as a matched-control for solid tumor somatic mutation profiling and for the detection of clinically-relevant variants in hematological malignancies. An increasing number of clinical studies demonstrate the value of detecting disease-specific biomarkers from blood for diagnosis, treatment monitoring, and cohort study recruitment.
Covaris Shearing Guide for 500 Base Pairs using the 96 oneTUBE-10 AFA Plate
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Covaris provides tools and technologies to improve pre-analytical sample preparation, enable novel drug formulations, and manage compounds in the drug discovery process.