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  • Bruno Vellutini 18:43 on 2014/07/01 Permalink
    Tags: , hox, , ,   

    Cloning genes that failed 

    Mmem wnt4a Nano hox1 Nano wnt4 Nano wnt7

    PIX

    Well, not that different… Membranipora gene worked somehow, a weak band.

    Set a ligation for Mmem wnt4a and Na hox1.

    03/07/14

    Heat shock transformation and plating.

     
  • Bruno Vellutini 17:01 on 2014/04/29 Permalink
    Tags: , , , hox, , ,   

    Cloning Ttra frizzled, Nano hox1, dlx, wnts 

    Need to clone the remaining frizzleds from Terebratalia and wnts from Novocrania. Set overnight regular PCR with:

    Nano hox1 Nano wnt4 Nano wnt7 Nano dlx Ttra fz5/8 Ttra fz7

    30/04/14

    20140430_175703

    Unfortunately hox1 and the wnts did not work. I should change something in the PCR. I extracted dlx and frizzleds and set a ligation for 4h at RT.

    Transformed and plated the 3 genes and incubated at 37 °C.

    01/05/14

    Pic colonies for colony PCR. Run gel and set bacteria to grow.

    20140501_150448

    02/05/14

    Extracted minipreps and set sequencing PCR.

     
  • Bruno Vellutini 21:32 on 2013/11/26 Permalink
    Tags: , hox, , , ,   

    Cloning Novocrania and Terebratalia again 

    Having the gene expression patterns of additional genes in Novocrania would be great to compare the body patterning with Terebratalia. I have selected some genes specific to apical, mantle, or pedicle lobe to try to map the expression of Terebratalia in Novocrania.

    • Apical lobe: wnt7, wnt8.
    • Mantle lobe: dlx, pax258, hox1.
    • Pedicle lobe: wnt4, wnt11.

    Also a couple of wnt genes that hadn’t worked in Terebratalia are also being cloned. I forgot to include Nano dlx in the order :/

    2013-11-26 21.29.18

    27/11/13

    Transformed and plated at 15h30. Put plates in incubator at 19h00.

    28/11/13

    Colony PCR and put cultures to grow. Wnt7 and Hox1 were not good so I plated them again with remaining bacteria.

    2013-11-28 18.31.05

    29/11/13

    Miniprep and sequencing PCR. Dropped sequences in facility.

    03/12/13

    BV275 Terebratalia transversa Wnt2 T7 ok + SP6
    BV276 Terebratalia transversa Wnt2 T7 ok + SP6
    BV277 Terebratalia transversa Wnt2 T7 ok + SP6
    BV278 Terebratalia transversa WntA T7 ok + SP6
    BV279 Terebratalia transversa WntA T7 ok + SP6
    BV280 Terebratalia transversa WntA T7 ok + SP6
    BV281 Novocrania anomala Wnt7 T7 ok
    BV282 Novocrania anomala Wnt11 T7 ok + SP6
    BV283 Novocrania anomala Wnt11 T7 ok T7
    BV284 Novocrania anomala Pax2/5/8 T7 ok + SP6

     
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