Extended Data Fig. 8: MNase protection assay to characterize binding patterns for P. furiosus histones incubated with the pCR7-long plasmid.
From: Histones direct site-specific CRISPR spacer acquisition in model archaeon
(a) Gel image of DNA fragments generated by micrococcal nuclease digestion of pCR7-long plasmid incubated with purified recombinant P. furiosus histones A and B. Colored boxes highlight major bands of protected DNA. The gel image is for a digest containing 100 units of MNase; a 500 unit digestion was also done and resulting DNA bands were much fainter. (b) DNA bands highlighted in A were sequenced and genome browser tracks were generated. Overlapping subtracks are color-coded according to the size of the DNA fragment (red: 30 bp +/- 5; golden yellow: 60 bp +/- 5; green: 90 bp +/- 5; blue 120 bp +/- 5). The full CRISPR7 and part of one repeat are shown, with the purple line indicating the position where integration would typically occur in vivo. The x-axis indicates the cumulative depth of read coverage. One replicate was done for each treatment (100 and 500 units MNase).
