User:Karmella Haynes/Notebook/Polycomb project/2011/04/03

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


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04/03/11

 * &#x2713; RT-PCR: test new primers
 * &#x2713; Repressed luc reactivation: expand HEK293 23;4;9 to 12 plates total (split each current one 1:3); dox induction (4x no dox, 4x 2-day, 4x 4-day)
 * &#x2713; Pc-TF cell cultures: split all cultures; 6-well plate dox induction (4 days) for 129-4, 154-2, 156-5, 157-1 (3 reps ea.)

RT-PCR > Test new RT-PCR primers on KAH126-1 (20 rxns total) > Primers: 1-4. HOXC4 50A-D (86, 87, 86, 85 bp) 5-8. IGFBP2 51A-D (117, 120, 113, 114 bp) 9-12. NFE2 52A-D (90, 88, 118, 120 bp) 13-16. NEUROG1 53A-D (105, 117, 92, 98 bp) 17-20. SOX1 54A-D (84, 80, 80, 102 bp)

> Template:
 * 1) KAH126-1 +dox (3/10/11) 1:10

--> 1 template mix: aliquot 19.0 μL --> Add 1.0 μL primers

--> PCR (96-well)
 * 95°C/ 3 min.
 * [95°C/ 15 sec., 58°C/ 15 sec., 72°C/ 15 sec.] x32
 * 72°C/ 3 min.
 * 4°C/ ∞



--> Conclusions:
 * 1) HOXC4: qPCR test A-D
 * 2) IGFBP2: qPCR test  A-D
 * 3) NFE2: failed
 * 4) NEUROG1: failed
 * 5) SOX1: failed

Next: Do qRT-PCR test with 1:10 dilution of 126-1 (+dox 3/10/11)


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