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Detection of a Cse4 mononucleosome at the CEN depends on the inclusion of anti-Cse4 antibody. MNase-treated chromatin prepared from strain SBY3 was immunoprecipitated with protein G Dynabeads in the presence (w/ Ab) and absence (w/o Ab) of anti-Cse4 antibody. Cse4-associated DNA was isolated and separated on a 1.5% agarose gel. Southern blotting was performed with a probe specific to the CEN3. M, radiolabeled 100-bp ladder marker; IN, input DNA; IP, the DNA bound to Cse4.
