User:Karmella Haynes/Notebook/Polycomb project/2010/05/03

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


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05/03/10

 * &#x2713; RT-PCR: 4-day dox Pc-ATF lines
 * &#x2713; Bact. cultures: FlpE (4)
 * &#x2713; Senescence assay cultures: expand/split confluent cultures
 * &#x2713; Pc-ATF negative ctrl lines: Pick colonies (96-well plates, 32 colonies/line)

RT-PCR

> Continued from 4/29/10: semi-qRT-PCR --> Use 1:100 dilution for mCh primer (10,000 might be too low) --> cDNA templates
 * 1) KAH126-1
 * 2) KAH126-1(+dox)
 * 3) KAH127-4
 * 4) KAH127-4(+dox)
 * 5) KAH128-8
 * 6) KAH128-8(+dox)
 * 7) KAH129-4
 * 8) KAH129-4(+dox)
 * 9) KAH130-2
 * 10) KAH130-2(+dox)
 * 11) KAH131-9
 * 12) KAH131-9(+dox)
 * 13) KAH132-8
 * 14) KAH132-8(+dox)
 * 15) KAH133-1
 * 16) KAH133-1(+dox)
 * 17) FTRx
 * 18) FTRx(+dox)

--> Primers; cDNA dilution
 * 1) p15INK(5); 1:1
 * 2) p16INK4a(7B); 1:1
 * 3) mCh f1/r1; 1:100
 * 4) GAPDH(21A); 1:10,000

--> Looks interesting; 126-1 appears to increase p16INK --> Redo GAPDH controls (21A and 21B primers) with 1:1000 dilution


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