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  • Bruno Vellutini 18:29 on 2015/04/29 Permalink
    Tags: , , , fgf8/17/18, , , ,   

    Membranipora cloning final genes 


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    Set PCR with:

    Mm bmp2/4 Mm otx1 Mm chordin Mm fgf8/17/18
    Mm nanos Mm piwi1 Mm piwi2 Mm vasa

    30/04/15

    Ran gel:

    2015-05-01 16.23.27

    Most important genes worked: bmp2/4, chordin and fgf8. piwi1 is bonus. Extract these and set a ligation for 3h. Transformed and plated around 18h.

    01/05/15

    Set colony PCR.

    2015-05-01 16.23.36

    Put colonies to grow.

    02/05/15

    Extracted minipreps, set sequencing PCR and dropped at seq facility.

    BV378 Membranipora membranacea Bmp2/4 T7 ok + SP6
    BV379 Membranipora membranacea Bmp2/4 T7 ok + SP6
    BV380 Membranipora membranacea Chordin T7 ok + SP6
    BV381 Membranipora membranacea Chordin T7 ok + SP6
    BV382 Membranipora membranacea Fgf8/17/18 T7 ok + SP6
    BV383 Membranipora membranacea Fgf8/17/18 T7 ok + SP6
    BV384 Membranipora membranacea Piwi1 T7 ok + SP6
    BV385 Membranipora membranacea Piwi1 T7 ok + SP6

     

     
  • Bruno Vellutini 10:30 on 2013/06/25 Permalink
    Tags: , fgf8/17/18, , , ,   


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    mamed2013_bcv

     
  • Bruno Vellutini 16:14 on 2012/11/05 Permalink
    Tags: , fgf8/17/18, , , , ,   

    Some cloning of Lineus and Priapulus 


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    Cloning of Pc gmcl, Lr caudal, Lr goosecoid, Lr foxq2, Lr fgf 8/17/18. Running the PCR with 60 °C for annealing, adjusted to primers Tm.

    06/11/12

    Running gel:

    I cut caudal and goosecoid bands. Extracted, ligated/transformed.

    09/11/12

    Colony PCR:

    10/11/12

    Minipreps for cloning:

    Colony T7
    cdx 6 BV202
    cdx 7 BV203
    cdx 8 BV204
    gsc 6 BV205
    gsc 7 BV206
    gsc 8 BV207

     
  • Bruno Vellutini 19:01 on 2012/10/29 Permalink
    Tags: , fgf8/17/18, , , , , ,   

    Lineus ruber first cloning 


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    Trying to clone 6 genes: six 3/6, caudal, goosecoid, foxa, foxq2, and fgf8/17/18. PCR was not so successful, only foxa and six3/6 worked:

    30/10/12

    Set ligation.

    31/10/12

    Transformed and plated.

    01/11/12

    Colony PCR for both genes:

    03/11/12

    Minipreps.

    Colony T7
    foxa 4 BV196
    foxa 5 BV197
    foxa 6 BV198
    six3/6 6 BV199
    six3/6 7 BV200
    six3/6 8 BV201

     
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