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  • Bruno Vellutini 18:28 on 2012/08/27 Permalink
    Tags: , , , nk6, , Prostheceraeus vittatus, ,   

    Probe synthesis for Terebratalia, Membranipora and Prostheceraeus 


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    Started a probe pcr with the following minipreps:

    Tt gli BV157 T7 3312,97
    Tt smo BV153 SP6 1154,37
    Tt nk6 BV173 SP6 1179,23
    Mm syt1a BV86 SP6 719,42
    Mm syt1b BV89 T7 942,87
    Pv syt1b BV189 T7 921,73
    Pv foxg BV191 SP6 1464,56
    Pv syt4a BV194 T7 1354,31

     
  • Bruno Vellutini 18:26 on 2012/08/27 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77

    It turns out that the cooling system was not properly working (water was not flowing through the tubes) and thus, temperature was high (room temperature). That’s why embryos were randomly failing to develop. Solution was to add more mqH2O to the cooling reservoir and flow began.

    I set a 4D with an embryo which looks ok and was sticking nicely to the slide, so it should not move, let’s see.

     
  • Bruno Vellutini 18:25 on 2012/08/17 Permalink
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    • 4D recording did not gastrulate.
    • Set a new one starting from 32 cell.
    • Fertilize one more 17/08/12 for late gastrula fixation.
     
  • Bruno Vellutini 20:57 on 2012/08/16 Permalink
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    • Spawning activated with EDTA at 12:30. Fixed for in situ and confocal at: 0-2h, 2-4h, 4-6h.
    • Spawning activated at 20:30 for fixing later stages.
    • Found out that pen-strep water is essential for 4D microscopy. Maybe it is a good idea to use it in normal cultures.
    • Set a recording that seems to be working.
     
  • Bruno Vellutini 18:22 on 2012/08/15 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77

    Fed colonies from 10/08 with Rhodomonas.

     
  • Bruno Vellutini 18:20 on 2012/08/11 Permalink
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    The way seems to be: spawn abundantly, use EDTA to activate for 1h, transfer embryos to a regular dish and wash a couple of times, split if necessary, wash in the next day, keep the concentration low.

     
  • Bruno Vellutini 18:17 on 2012/08/09 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77

    Made a spawning which yielded a huge amount of eggs. I divided in 4 samples:

    1. Agar-coated 7 °C / 15 °C
    2. Regular dish 7 °C / 15 °C

    Also added Rhodomonas to all 3 culture dishes with cyphonautes.

     
  • Bruno Vellutini 15:57 on 2012/08/09 Permalink
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    Probe synthesis of wnt genes 


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    Sample ID  ng/ul
    Tt wnt1    1959,05
    Tt wnt5    939,65
    Tt wnt6    799,08
    Tt wnt9    519,62
    Tt wnt10 1736,08
    Tt wnt16 2439,94

     
  • Bruno Vellutini 11:47 on 2012/08/07 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77
    • Pipeting the embryos before the EDTA treatment into gelatin coated dish did not work so well. Seems to be a lower ratio of cleaving embryos.
    • At least one more well developed larva fed on Rhodomonas. I think it is worth trying.
    • Changed the dish for the 29/07/12 dish because larvae were getting stuck in the thin layer of “stuff” growing in the bottom. Some stuck at the bottom of the new clean dish, though…
     
  • Bruno Vellutini 18:13 on 2012/08/06 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77

    Tried to feed early cyphonautes (8 day old) with Rhodomonas. One larva captured the particle (into the atrium), but it remained stuck in the mouth region. Maybe it is not ready yet? After a couple of minutes the larva ejected the algae.

     
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