[PRECELLYS] Purification of High Molecular Weight DNA from Cannabis sativa

Long-read sequencing techniques such as Oxford Nanopore and PacBio require very long and very pure DNA. Because small fragments are preferentially sequenced, it is important to limit their presence to increase downstream contiguity. Plant cells are encased in a defensive cell wall and are frequently 80% vacuole. Isolation of nuclei removes the large fraction of nucleases and other hydrolytic components that can attack DNA upon lysis.

Cannabis tissues are rich in polyphenols, and so the use of reducing agents such as PVP (Polyvinylpyrrolidone) and BME (ß-mercaptoethanol) are essential. Because important genes, such as the cannabinoid synthases, are highly duplicated and separated by large repetitive regions, long-read sequencing is necessary to resolve their structure with high fidelity.

Here the Precellys Evolution homogenizer has been used to isolate nuclei from Cannabis sativa plant cells and obtain high molecular weight DNA, compatible with NGS long-read sequencing techniques .