User:Karmella Haynes/Notebook/Polycomb project/2010/06/08

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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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06/08/10

 * &#x2713; ChIP: pull-down, wash non-specific binding & elution, DNA purification
 * &#x2713; ChIP Western: quality control analysis Western blot
 * &#x2713; Growth assay: add fresh 1 μg/mL dox to all dox+ samples

ChIP: quality control analysis Western blot > Did chromatin-ab + bead binding for 4 hours > Pronase treatment overnight (continue tomorrow) > Western blot samples: > Use 10-well gel (loading volume = 25 μL) > Electroblot: 1 hr. 15 min.
 * 1) mouse α-PolII ChIP Supernatant
 * 2) mouse α-PolII ChIP (eluted)
 * 3) mouse α-IgG ChIP Supernatant
 * 4) mouse α-IgG ChIP (eluted)
 * 5) Input

--> Primary: mouse α-PolII (ab817), 1:1000, 5 mL

6/09/10 --> Secondary: donkey α-mouse IgG-HRP, 1:5,000 (R.T./ 1 hr.) --> PolII = 217 kD --> Pull-down unsuccessful. Try again using more chromatin and more beads.


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