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  • Bruno Vellutini 18:36 on 2014/06/06 Permalink
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    Cloning Membranipora Wnts 


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    Set PCR.

    07/06/14

    Gel, extraction, ligation, transform.

    09/06/14

    Set colony PCR overnight.

    10/06/14

    Ran gel and put colonies to grow.
    2014-06-10 12.25.20

    11/06/14

    Extracted minipreps and did sequencing PCR. Left samples at the sequencing facility.

     
  • Bruno Vellutini 16:03 on 2014/06/06 Permalink
    Tags: , , , , , ,   

    Brachiopod in situ 


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    In situ for additional Terebratalia frizzleds and repeating some Novocrania for better pictures.

    Nano engrailed wnt1 wnt5 dlx pax258
    Ttra frizzled 5/8 frizzled 7 dcr1b tdr1 ago

    Started in situ with standard procedures. 8 min protk for Novocrania and 10 min for Terebratalia.

    07/06/14

    Added probes, embryos look good.

    09/06/14

    Washing day. Everything went as usual and put antibody around 18h.

    10/06/14

    Washed with PBT several times berween 10-15 min. Then PTw 30 min washes. Started developing.

    Stopped Nano dlx after 2h30min because it was done, as well as Ttra fz5/8. The rest continued developing. fz7 got pinkish really fast

    11/06/14

    fz7 and ago got really dark. I don’t know if it is background or signal, even though it looks like there is real signal within the background. I stopped them as well as engrailed and pax258, which look better than the last in situ.

    12/06/14

    Stopped wnt1, wnt5 dcr1b, tdr1. Novocrania samples are looking dark ugly already…

    13/06/14

    Trying to clear out some of the background from fz7 and ago… letting in ethanol for many hours.

     
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