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:
1.
Samples were phenol:chloroform extracted and EtOH precipitated:
- Added equal volume of phenol:chloroform:IAA, vortexed, spun @ 12,500g, 5mins, 4C.
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Transferred aqueous phase to clean tube.
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Added equal volume of chloroform, vortexed, spun @ 12,500g, 5mins, 4C.
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Transferred aqueous phase to clean tube.
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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.