PcG perturbation experiment planning
- Cavalli lab (Gonzalez et al) do RNAi based visual screen of RNAi knockdown, look at Pc.
- distributions of Pc fluorescence across cells in Pc knockdown is pretty tight — the remaining variability could be background auto-fluorescence. This looks promising.
- recommend dsRNA targeting th and rho as transfection controls, these produce cell death and large nuclear size.
- flyRNAi protocol: http://www.flyrnai.org/DRSC-PRS.html (from Drosophila RNAi screening center at Harvard Medical School).
- see also http://www.genomernai.org/About,
- do I just look up tested-primer sequences (with T7 additions) for my targets and use these to amplify from my in-house genomic DNA?
- or can I order DNA template from the DRSC?
- Nice PSC Su(Z)2 deletion: Su(Z)2-1.b8
- could order corresponding fly line to make late-stage embryos as well: 24467. Deletion over eve-LacZ marked balancer SM6 (Cy).
- should contact Perrimon lab.
- more Sneider’s medium (for culture of deletion line and for RNAi protocols).
- Fly stock: 24467
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