ChIP-qPCR was performed essentially as previously described (Graf et al., 2017 (link)). For protein A ChIP, IgG Sepharose 6 Fast Flow beads (GE Healthcare) were used. For Myc ChIP, nProtein A Sepharose 4 Fast Flow beads (GE Healthcare) were used; after preclearing, 9 µl anti-c-Myc Monoclonal antibody (Clontech/Takara) were added to each sample. qPCR was performed using a LightCycler 480 II (Roche) and SYBR-Green (Thermo Scientific) detection with an annealing temperature of 60°C (40 cycles). Primers are listed in Supplementary file 1C . Measured Cq values were corrected to input and graphs were created using R. All strains were propagated for ~50 generations before ChIP-qPCR analysis.
Nprotein a sepharose 4 fast flow beads
Manufactured by GE Healthcare
NProtein A Sepharose 4 Fast Flow beads are a chromatography resin used for the purification of antibodies and other proteins that bind to Protein A. The beads are made of cross-linked agarose and have a high capacity for Protein A ligand. They are designed for fast protein purification workflows.
Automatically generated - may contain errors
2 protocols using nprotein a sepharose 4 fast flow beads
1
ChIP-qPCR for Protein-DNA Interactions
2
ChIP-qPCR Protocol for Protein A and Myc
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ChIP-qPCR was performed essentially as previously described (Graf et al., 2017) (link). For protein A ChIP, IgG Sepharose 6 Fast Flow beads (GE Healthcare) were used. For Myc ChIP, nProtein A Sepharose 4 Fast Flow beads (GE Healthcare) were used; after preclearing, 9 µl anti-c-Myc Monoclonal antibody (Clontech/Takara) were added to each sample. qPCR was performed using a LightCycler 480 II (Roche) and SYBR-Green (Thermo Scientific) detection with an annealing temperature of 60ºC (40 cycles). Primers are listed in Supplementary Table S3. Measured Cq values were corrected to input and graphs were created using R. All strains were propagated for ~50 generations before ChIP-qPCR analysis.
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