Protocol to isolate nuclei from Chlamydomonas reinhardtii for ATAC sequencing

Area of Science:

• Molecular Biology
• Genomics
• Plant Science

Background:

• High-resolution chromatin accessibility mapping via assay for transposase-accessible chromatin using sequencing (ATAC-seq) requires intact nuclei.
• Current methods for nuclei isolation in Chlamydomonas reinhardtii can be time-consuming and may affect sample quality.

Purpose of the Study:

• To present an optimized, tag-free protocol for the efficient isolation of high-quality nuclei from Chlamydomonas reinhardtii.
• To enable downstream ATAC-seq analysis for chromatin accessibility studies in this model alga.

Main Methods:

• Developed a novel protocol for tag-free nuclei isolation from both cell-walled and cell-wall-deficient Chlamydomonas reinhardtii strains.
• Included detailed steps for nuclei isolation, quantification, and preparation for ATAC-seq.
• Optimized the protocol to reduce the overall time for nuclei isolation and quantification.

Main Results:

• Successfully isolated intact nuclei suitable for ATAC-seq from diverse Chlamydomonas reinhardtii strains.
• Demonstrated the efficiency and speed of the developed protocol compared to existing methods.
• Ensured the quality of isolated nuclei for reliable downstream genomic analyses.

Conclusions:

• The presented protocol provides a streamlined and effective method for nuclei isolation in Chlamydomonas reinhardtii.
• This facilitates high-throughput chromatin accessibility studies using ATAC-seq in this important model organism.
• Enables deeper insights into gene regulation and epigenetic mechanisms in green algae.