Covaris, Inc.

Hans-Ulrich Thomann

James A Laugharn, Jr

Hamid Khoja

cfDNA reference material, for example, for use in verifying the accuracy and effectiveness of a diagnostic test, is generated from chromatin which may be sourced from whole cells. The chromatin may be treated with formaldehyde to form crosslinks between DNA and histones, for example, while the chromatin is contained within nuclear and/or plasma membranes. The fixed chromatin may be sheared by acoustic energy, which may also be used to lyse cell membranes within which the fixed chromatin may be contained. The sheared chromatin may be treated with an enzyme, such as micrococcal nuclease, to digest chromatin in linker regions of the DNA between nucleosomes and generate nucleosome material for use as cfDNA reference material.

Generation of cfDNA reference material

Focused acoustic treatment of samples may be used to differentially shear different fragment lengths of DNA or other nucleic acid portions of a sample. Relatively larger fragment lengths may be sheared while smaller fragment lengths are unaffected by the focused acoustic based shearing.

Differential shearing of nucleic acids

Focused acoustic treatment of samples including a target biomolecule, such as cfDNA, may aid in the recovery of the biomolecule from a sample. cfDNA in a sample, whether chemically stabilized or not, may be linked or otherwise bound to histones or other proteins, e.g., by hydrogen bonds of histones or chaperone proteins to DNA and/or covalent crosslinks of such proteins to the DNA. Focused acoustic energy may remove or disrupt such links, aiding in isolation of the cfDNA from the sample.

Extraction of cfDNA from biological samples

Systems and methods for collecting blood and other liquid samples. A sample holder may include a cap and porous element arranged to receive a blood or other liquid sample. The porous element may be stored in a sealed container, e.g., immediately after blood sample collection, and the sample dried while sealed in the container.

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Stow, MA, United States of America

Hans-Ulrich Thomann