User:Karmella Haynes/Notebook/Polycomb project/2010/11/30

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11/30/10

  • ✓ ChIP: sonicate U2OS FTRx chromatin; H3K27me3 IP overnight



Sonication
> U2OS FTRx cells

  1. Double cross-linked "dx" (DMA, CH2O)
  2. Single cross-linked "fx" (CH2O)


IP: antibody binding

  1. FTRx dx, H3K27me3 07-449
  2. FTRx dx, rabbit IgG
  3. FTRx fx, H3K27me3 07-449
  4. FTRx fx, rabbit IgG

Note: Out of rabbit IgG! Use rabbit α-myc

Reagent 1, 3 2, 4
α-H3K27me3 07-449 2.0 ---
rabbit α-myc --- 2.0
Chromatin 500.0 500.0

--> Rotate at 4°C/ overnight