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  • Bruno Vellutini 18:06 on 2012/04/18 Permalink
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    Rehydrate the whole egg mass to see if it was easy to take the embryos out. Kind of easy to dissect since jelly is no longer sticky and membrane is disrupted when taking the bundle off the matrix. However, embryos are slightly deformed due to dehydration.

     
  • Bruno Vellutini 18:00 on 2012/04/18 Permalink
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    Cloning Membranipora and Terebratalia piwi, nanos, vasa, hedgehog, smoothened, runx 


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    Running a PCR overnight to clone the following genes:

    Mm: Piwi1, Vasa, Runx
    Tt: Hedgehog, Smoothened, Piwia, Piwib, Vasa, Nanos.

    First time cloning Terebratalia genes.

    19/04/12

    Membranipora bands were ok and were purified directly (and ligated), but Terebratalia were too faint:

    For this reason I ran a re-PCR for Terebratalia genes.

    20/04/12

    For the re-PCR I diluted the PCR product of the first reaction 1:20 and used as a template for a regular PCR (without changing any parameters). The bands this time were stronger, except for Piwia. Since unwanted bands appeared I had to cut the gel, extract, and purify the DNA.

    Ligation was also set for these genes.

    21/04/12

    Heat shock transformation for all genes.

     
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