MeDIP - SB/WB Fragmented gDNA (continued from yesterday)

2010
Miscellaneous
Author

Sam White

Published

July 2, 2010

Continued MeDIP process from yesterday. Protein A/G beads were pelleted XXXXXXXXX, supe transferred to clean tube. Beads were washed 3x in the following fashion, each wash saved to retain unmethylated DNA:

Samples were phenol:chloroform extracted and EtOH precipitated:

  1. Added equal volume of phenol:chloroform:IAA, vortexed, spun @ 12,500g, 5mins, 4C.

  2. Transferred aqueous phase to clean tube.

  3. Added equal volume of chloroform, vortexed, spun @ 12,500g, 5mins, 4C.

  4. Transferred aqueous phase to clean tube.

  5. Added 0.1 vols 3M NaOAc (pH=5.2), 2.5 vols of 100% EtOH, mixed and stored @ -20C over the weekend.

Will finish precipitation next week and quantify recovery.