A. pG5ML plasmid was relaxed by Topoisomerase I and incubated in oocyte extract for up to 5 hours as indicated. Reactions were stopped at indicated time, deproteinized and the DNA was run on an agarose gel and stained with EtBr. Supercoiled and relaxed positions are indicated.
B. pG5ML plasmid was relaxed by Topoisomerase I and incubated in egg extract for up to 1.5 hours as indicated. Reactions were stopped at indicated time, deproteinized and the DNA was run on an agarose gel and stained with EtBr. Supercoiled and relaxed positions are indicated
C. Plasmid DNA was incubated in oocyte extract for 2.5 hours and reactions were treated with 1U of micrococcal nuclease (MNase) for 0, 5, 10, 15, or 20 minutes. Reactions were deproteinized and the DNA was run on an agarose gel and stained with EtBr. Marker positions are indicated in basepair length. Nucleosomes are 146bp to 200bp fragments.
D. Plasmid DNA was incubated in egg extract for 2.5 hours and reactions were treated with 1U of micrococcal nuclease (MNase) for 0, 5, 10, 15, or 20 minutes. Reactions were deproteinized and the DNA was run on an agarose gel and stained with EtBr. Marker positions are indicated in basepair length. Nucleosomes are 146bp to 200bp fragments.