User:Karmella Haynes/Notebook/Polycomb project/2010/05/09

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05/09/10

  • ✓ Pc-ATF neg ctrl lines: screen for RFP; choose 6 col's per line
  • ✓ Misc cell culture: humidifying water was dried up in the incubator; lost 126-4; split others (2x 10cm plates, one for frozen stocks)
  • ✓ KAH126-4: thaw new stock (last one!)



Pc-ATF neg ctrl lines RFP screen
> 96-well plates, 32 wells per line
> Screen for nuclear dox-inducible expression of RFP; RFP+ in dox+ and RFP- in dox-
> Key (dox+/dox-)
--> - = no expression
--> + = weak/ non-uniform expression
--> ++ = low or medium uniform expression
--> +++ = very strong expression (uniform or non-uniform)
--> ? = odd morphology/ strange RFP localization


Line 1 2 3 4 5 6
KAH154 ++/- ++/- ++/- +++/+ ++/- ++/-
KAH155 ++/+ ++/- ++/+ ++/+ +++/+ ++/-
KAH156 +++/- +++/+ ++/- ++/- ++/- +++/+
KAH157 ++/- ++/- ++/- ++/- ++/- ++/-
KAH158 ++/- ++/- ++/- ++/- ++/- +/-
KAH159 ++?/- ++?/- ++?/- ++?/- +/- +/-

--> Note: KAH159 clones with ++ expression show a strange single bright dot of RFP, plus medium level RFP expression throughout the nucleus. Nuclear protein aggregate?

--> Tomorrow: transfer dox- cells to 24-well plates



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