Introduction

The invention relates to a sequencing method, able to detect chromatin changes, specifically heterocromatin. Alterations in heterochromatin are associated with developmental defects and cancer, while its proper conformation is a hallmark of healthy cells. As such, reliable methods to characterize heterochromatin and its alterations are essential for the biomedical scientific community.

Technical features

Cromatin, found in the cell nucleus has a complex structure, and in particular heterochromatin, a compact derivative, is highly specific to the nucleus. Alterations in the heterochromatin structure or localization is a hallmark of disease and cancer, however the known procedures to analyzed the heterocromatin levels, require identifying specific targets, whose hard accessibility and distribution can be a limitation in the efficiency of such methods. This invention includes a protocol for chromatin fractionation, to facilitate the mapping of the heterocromatin accessibility. The procedure can be applied to a small number of primary cells and does not involve chemical modification which may cause high background noise during sequencing. The protocol requires only about 3 hours of bench work.

Possible Applications

• Alteration detection in chromatin structure and functions;
• Diagnosis of pathologies that alter genome structure, as cancer or laminopathies;
• DNA sequencing.

Advantages

• Method applicable on various types of primary cells of various types;
• No chemical modification of chromatin;
• There is no need for antibodies and therefore greater flexibility in terms of experimental design;;
• Reproducible and quick (3 hours);
• Reduced number of initial cells.