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  • Bruno Vellutini 00:12 on 2014/05/13 Permalink
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    Confocal with Membranipora propidium iodide and phalloidin far red 

    Round of confocal with wild type Membranipora embryos. I mistakenly stained with propidium iodide instead of sytox green, but well. They were mounted in SlowFade to see if this is better than glycerol. However, it seems that it is incompatible with propidium and the nuclear staining became a general cytoplasm staining.. Many interesting scans of early and late gastrulation.

    MAX_Mmem pi-phallred.lif - Series012

     
  • Bruno Vellutini 00:06 on 2014/05/13 Permalink
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    Washed multiple times Lineus longissimus juveniles for RNA extraction. Put them into a RNA tube and wahsed and centrifuged more than 3 times with uvfiltered sea water. Then snapfreeze. They were around 40 days old.

     
  • Bruno Vellutini 16:12 on 2014/05/08 Permalink
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    Washed Lineus longissimus larvae. Many larvae in the more advanced jar at 19 °C had already settled. I removed the swimming larvae by filtering the water until there was only a finger of water left. Then I rinsed the bottom to release the juveniles and transferred everything to a glass bowl.

    Juveniles look happy. I washed and cleaned the bowl as much as possible, but the juveniles stick to the bottom and to the pipets, so it is a bit hard. I transferred most of the worms to a clean bowl and kept the dirty as well in the incubator at 14 °C. I’ll separate many for RNAlater, a few for immunos and the rest to raise until adulthood.

    The other jar in fritzen room does not look so good. I’ll have a look at it next week. The jar from the incubator has many larvae, but they have the least advanced rudiment worm inside.

    I forgot to add food. I will add tomorrow.

     
  • Bruno Vellutini 10:11 on 2014/05/08 Permalink
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    Probe synthesis Ttra frizzleds Nano dlx and more 

    Gene Miniprep Antisense Enzyme ng/µL
    Nano dlx BV285 SP6  159
    Ttra fz5-8 BV288 SP6  366
    Ttra fz7 BV291 SP6  2439
    Ttra dicer1b BV162 SP6  1867
    Ttra tudor1 BV167 SP6  1076
    Ttra argonaute BV168 SP6  323

    Started probe PCR.

    2014-06-05 17.42.43

    09/05/14

    Began probe synthesis at 10:30.

    10/05/14

    Done with probe synthesis, see values above.

     
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