A good strategy to optimize MNase concentration for bacterial Ribo-seq?

16 April 2017 0 9K Report

Micrococcal nuclease(MNase), is a nuclease of choice for performing bacterial ribosome footprints. The literature vouches for MNase to be efficient with low cleavage of rRNA.

During my use of the enzyme i have found even low levels of MNase(8 kunitz units/A260) ribosome, i find significant cleavage of the 23S rRNA. Does anyone have experience with optimizing the use of MNase with ribosomes for preparation footprints with minimal degradation of rRNA without compromising the yield of footprints?

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