User:Karmella Haynes/Notebook/Polycomb project/2010/07/24

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07/24/10

 * &#x2713; Transfections: Test Gal4-ATF's on KAH142 H3me reporter clones

Fugene transfection > 12-well plate 1-4. KAH142-1 5-8. KAH142-3 9-12. KAH142-5

> Master mixes (3x mix, 4 mixes): 5.4 μL Fugene + 54.6 μL Opti-MEM --> R.T/ 5 min. > Add 3x vol DNA --> R.T./ 20 min. > Add 20 μL complexes to each well (1 ml med. each); Grow cells at 37&deg;C overnight

7/25/10 Results:
 * Background YFP expression: two or three cells for KAH142-5, none for KAH142-3, KAH142-1 control well was contaminated
 * KAH54/pcVN (Gal4DB-VP64): YFP+ cells in KAH142-3
 * KAH59/pcVN (Gal4DB-mCh-mCh-VP64): RFP+/YFP+ cells in KAH142-3 and KAH142-5
 * KAH60/pcVN (Gal4DB-mCh-VP64): strange result; RFP+/YFP- or RFP-/YFP+ cells

Conclusions:
 * Keep KAH142-3 and 142-5 for frozen stocks; use 142-3 for further experimentation
 * Use KAH59/pcVN to screen other H3me Reporter clones


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