Frog histone reconstitute CAD
Ovis aries/Saccharomyces cerevisiae mixed DNA library
Two plasmid DNAs were employed: pBLG (BLG) comprised 10,841 bp of ovine DNA containing the beta-lactoglobulin gene and 2,020 bp of plasmid vector; p13 (YRO) comprised 13,626 bp of Saccharomyces cerevisiae DNA (chrXIV:243,179-256,806 ((SacCer_Apr2011/sacCer3) assembly)) containing a late-firing replication origin (ARS1413 (~ 250,600-251,220) and 6683 bp of plasmid vector. A mixture of equal weights of linearised plasmids containing BLG and YRO were reconstituted with recombinant Xenopus laevis core histones by salt gradient dialysis. Nucleosomal DNA was prepared from these reconstitutes using CAD nuclease. The resulting ~146 bp mononucleosome DNA fragments were purified after electrophoresis.