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"I think the field is pre-cleared.
The supernatant was saved after centrifugation and pre-cleared with M-280 Streptavidin Dynabeads (Life Technologies).
Pre-cleared lysates were incubated with anti-Flag beads (Sigma-Aldrich) with gentle rocking overnight at 4 °C.
Pre-cleared lysates were subjected to immunoprecipitation with indicated antibodies.
The sonicated lysates were pre-cleared using protein G sepharose.
10% of the pre-cleared lysate was set aside as input control.
The supernatant was collected by centrifugation and pre-cleared with protein A agarose beads (Millipore).
Specific antibody was then added to the pre-cleared supernatant and mixed at 4°C overnight.
Embryo lysate was diluted 1∶3 in ESB and pre-cleared with Protein G/A beads.
Pre-cleared soluble chromatin was incubated with 5 µg of antibodies.
The pre-cleared extract was then applied to the Sepharose-RNA column four times.
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