User:Karmella Haynes/Notebook/Polycomb project/2010/05/06

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05/06/10

 * &#x2713; Minipreps: FlpE (4)
 * &#x2713; X-linked chromatin prep & sonication: Dox-induced samples from 3/09/10 and 3/14/10; practice run for U2OS plain only (with Qingqing)
 * &#x2713; FlpE: cultures (4); streak 100μg/mL plate with clone #3

Minipreps > Check with E, P, and E/P digests

15 μL --> 37°C/ ~15 min.

--> Conclusion: Success! Combine 4 preps (300μL total) for DNA clean-up (tomorrow) --> Streak clone #3 (saved toothpick) onto 100 μg/mL Amp plate. --> Inoculate 4 5 mL cultures and 1 2 mL culture (for glycerol stock) w/ 2 μL left-over culture #3.

X-linked chromatin prep & sonication > Full sample set is numbered as follows:
 * 1) KAH126-1 (3/14/10)
 * 2) KAH127-4 (3/09/10)
 * 3) KAH128-8 (3/09/10)
 * 4) KAH129-4 (3/14/10)
 * 5) KAH130-2 (3/09/10)
 * 6) KAH132-8 (3/09/10)
 * 7) U2OS plain (3/14/10)

> Today, do chromatin prep and sonication for U2OS plain only ...Oops, grabbed #6 instead of #7 (5/21/10) > Completion time: ~2 hours for chromatin prep, ~2 hours per sample for sonication > No more than two samples should be done per day, since sonication must be done one sample at a time. > Finish at DNA quality control step. Pronase A treatment of 100 μL sample (overnight). Flash-freeze remaining ~6 mL and store at -80°C.


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