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  • Bruno Vellutini 16:54 on 2012/08/02 Permalink
    Tags: apical organ, , , , , , orthopedia,   

    RACE PCR for Pc gmcl, Mm nk2.1, otp, bruno 


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    Setting the first round of RACE PCR for:

    Pc gmcl1 F1/F2, Pc gmcl2 R1/R2, Mm nk2.1 F1/F2, Mm otp R1/R2, Mm celf2 F1/F2.

    Letting overnight.

    03/08/12

    Extraction and ligation.

    04/08/12

    Transformation and plating.

    06/08/12

    Colony PCR

    13/08/12

    Sequencing reaction for Pc gmcl genes.

    BV177 GMCL1 T7
    BV178 GMCL1 T7
    BV179 GMCL1 T7
    BV180 GMCL1 T7
    BV181 GMCL1 T7
    BV182 GMCL2 T7
    BV183 GMCL2 T7

     
  • Bruno Vellutini 16:51 on 2012/08/02 Permalink
    Tags:   


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    A relative small amounts of embryos of the 28/07 sample is growing into the late gastrula stage. I am transferring them to a clean 35 mm everyday.

    I tried a different strategy for increasing the ratio of developing embryos. Instead of collecting the spawning eggs from the bottom and washing many times and then putting them in EDTA water I let them in the bowl after adding 20 µL of EDTA, in the dark for ~2 h at 7 °C. I did not notice a higher amount of activated eggs, it seems that less eggs were activated. After this period I transferred them to a dish and washed. Lets see how they develop.

     
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