User:Karmella Haynes/Notebook/Polycomb project/2010/09/28

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09/28/10

 * &#x2713; RT-PCR: biological replicates for 4-day dox induction of Pc-ATF subset

RT-PCR > Pc-ATF biological replicates (30 total) --> Templates (6/15, 6/18, 6/23/10) --> Primers
 * 1) KAH126-1 -dox
 * 2) KAH126-1 +dox
 * 3) KAH126-3 -dox
 * 4) KAH126-3 +dox
 * 5) KAH132-8 -dox
 * 6) KAH132-8 +dox
 * 7) KAH154-2 -dox
 * 8) KAH154-2 +dox
 * 9) FTRx -dox
 * 10) FTRx +dox
 * 1) p16INK 7C (94 bp), (0.5 μL 1:1 cDNA) (30 rxns)
 * 2) GAPDH 21A (100 bp), (0.5 μL 1:1000 cDNA) (30 rxns)
 * 3) MMP12 31A (127 bp), (5.0 μL 1:100 cDNA) (30 rxns)
 * 4) mCh (100 bp), (1.0 μL 1:1000 cDNA) (30 rxns)
 * 5) THPO 33A (103 bp)
 * 6) TNF 34A (119 bp)

--> 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: Looks like MMP12 is induced just by the KAH126 proteins. Cool! Repeat MMP12 and p16 with 2x more template


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