Blocking Assay:Article Title: The DNA methylation landscape of human melanoma
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-tagged MBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10x NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated E. coli JM110 genomic DNA) for 20 min at 4°C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4°C on a rotating platform. .. To allow selective collection of MIRA-captured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 hours at 4°C on a rotating platform.
Article Title: The DNA methylation landscape of human melanoma.
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-taggedMBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10× NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated Escherichia coli JM110 genomic DNA) for 20 min at 4 °C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4 °C on a rotating platform. .. To allow selective collection of MIRAcaptured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 h at 4 °C on a rotating platform.
Sonication:Article Title: The DNA methylation landscape of human melanoma
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-tagged MBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10x NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated E. coli JM110 genomic DNA) for 20 min at 4°C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4°C on a rotating platform. .. To allow selective collection of MIRA-captured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 hours at 4°C on a rotating platform.
Article Title: The DNA methylation landscape of human melanoma.
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-taggedMBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10× NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated Escherichia coli JM110 genomic DNA) for 20 min at 4 °C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4 °C on a rotating platform. .. To allow selective collection of MIRAcaptured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 h at 4 °C on a rotating platform.
Incubation:Article Title: The DNA methylation landscape of human melanoma
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-tagged MBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10x NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated E. coli JM110 genomic DNA) for 20 min at 4°C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4°C on a rotating platform. .. To allow selective collection of MIRA-captured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 hours at 4°C on a rotating platform.
Article Title: The DNA methylation landscape of human melanoma.
Article Snippet: .. In brief, 1.5 μg of GST-tagged MBD2b protein and 1 μg of His-taggedMBD3L1 protein were pre-incubated in 355 μl of blocking buffer (40 μl of 10× NEBuffer 2, 1 μl of 10% Triton X-100, and 0.5 μg of sonicated Escherichia coli JM110 genomic DNA) for 20 min at 4 °C on a rotating platform and added to each ligated DNA sample, and then the 400 μl of the MIRA reaction mixture was incubated overnight at 4 °C on a rotating platform. .. To allow selective collection of MIRAcaptured methyl-CpG-rich DNA fragments, 5 μl of MagneGST magnetic beads (Promega, Madison, WI), pre-blocked with blocking buffer, were added to the MIRA reaction mixture and further incubated for 2 h at 4 °C on a rotating platform.
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