User:Karmella Haynes/Notebook/Polycomb project/2010/09/19

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


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09/19/10

 * &#x2713; RT-PCR: EZH2 kd cDNA, NS candidate genes; re-do MMP12, THPO, TNF in NS cDNA set (1:10 dilutions)

RT-PCR > EZH kd set --> Templates --> Primers
 * 1) n.t. siRNA 4-day, 11/03/09
 * 2) EZH2 kd 4-day, 11/03/09
 * 3) n.t. siRNA 8-day, 11/03/09 (?)
 * 4) EZH2 kd 8-day, 11/03/09 (?)
 * 1) p16INK 7C (94 bp) (4 rxns)
 * 2) MMP12 31A (127 bp) (4 rxns)
 * 3) THPO 33A (103 bp) (4 rxns)
 * 4) TNF 34A (119 bp) (4 rxns)

> NS cDNA set --> Templates (1:10 dilutions) --> Primers
 * 1) MMP12 31A (127 bp) (16 rxns)
 * 2) THPO 33A (103 bp) (16 rxns)
 * 3) TNF 34A (119 bp) (16 rxns)

--> PCR (96-well)
 * 95°C/ 3 min.
 * [95°C/ 30 sec., 57°C/ 30 sec., 72°C/ 30 sec.] x32
 * 72°C/ 3 min.
 * 4°C/ ∞



> Conclusions:
 * EZH2 kd: looks like genes are de-repressed after EZH2 kd, but p16INK is very weak. Do PCR again on full set of EZH2 kd's
 * Pc-ATF repeats: choose best trial for figure (vs. previous trial with more template)


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