User:Karmella Haynes/Notebook/Polycomb project/2010/02/20

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


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02/20/10

 * &#x2713; RNA/ cDNA prep: 4-day dox Pc-ATF cultures (plate set A)
 * &#x2713; Pc-ATF RT-PCR cultures: split plate set B into two duplicates; start new plate for KAH130-2
 * Cytology: seed 24-well cover slip plate for H3K27me/ H3K9me staining Very sparse cell growth. Do over on Tuesday
 * &#x2713; Cultures: Split confluent Pc-ATF lines 1:10 (10 cm plates)

RNA Preps/ cDNA synth.

> RNA Preps --> Use 1 mL TriZol/ well (6-well dishes) --> Resuspend RNA pellet in 10 μL THE RNA Storage Solution

> oligo(dT) Primer annealing

> cDNA synthesis mix --> 8 reactions total

--> Add 10 μL mix to each annealing rxn. --> 50°C/ 50 min., 80°C/ 5 min., ice --> Add 1.0 μL RNase H, 37°C/ 20 min. --> Store at -20°C


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