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The effect of micrococcal nuclease digestion on nucleosome positioning data.


ABSTRACT: Eukaryotic genomes are packed into chromatin, whose basic repeating unit is the nucleosome. Nucleosome positioning is a widely researched area. A common experimental procedure to determine nucleosome positions involves the use of micrococcal nuclease (MNase). Here, we show that the cutting preference of MNase in combination with size selection generates a sequence-dependent bias in the resulting fragments. This strongly affects nucleosome positioning data and especially sequence-dependent models for nucleosome positioning. As a consequence we see a need to re-evaluate whether the DNA sequence is a major determinant of nucleosome positioning in vivo. More generally, our results show that data generated after MNase digestion of chromatin requires a matched control experiment in order to determine nucleosome positions.

SUBMITTER: Chung HR 

PROVIDER: S-EPMC3012088 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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The effect of micrococcal nuclease digestion on nucleosome positioning data.

Chung Ho-Ryun HR   Dunkel Ilona I   Heise Franziska F   Linke Christian C   Krobitsch Sylvia S   Ehrenhofer-Murray Ann E AE   Sperling Silke R SR   Vingron Martin M  

PloS one 20101229 12


Eukaryotic genomes are packed into chromatin, whose basic repeating unit is the nucleosome. Nucleosome positioning is a widely researched area. A common experimental procedure to determine nucleosome positions involves the use of micrococcal nuclease (MNase). Here, we show that the cutting preference of MNase in combination with size selection generates a sequence-dependent bias in the resulting fragments. This strongly affects nucleosome positioning data and especially sequence-dependent models  ...[more]

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