Team:UT Dallas/NotebookWeek6

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         <li class = "active"><a href="https://2011.igem.org/Team:UT_Dallas/Notebook"><font size="3" face="verdana">Notebook</a></font></li>
         <li class = "active"><a href="https://2011.igem.org/Team:UT_Dallas/Notebook"><font size="3" face="verdana">Notebook</a></font></li>
         <li><a href="https://2011.igem.org/Team:UT_Dallas/HumanPractices"><font size="3" face="verdana">Human Practices</a></font></li>
         <li><a href="https://2011.igem.org/Team:UT_Dallas/HumanPractices"><font size="3" face="verdana">Human Practices</a></font></li>
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         <li><a href="https://2011.igem.org/Team:UT_Dallas/Gallery"><font size="3" face="verdana">Gallery</a></font></li>
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         <li><a href="https://2011.igem.org/Team:UT_Dallas/Safety"><font size="3" face="verdana">Safety</a></font></li>
          
          
       </ul>
       </ul>
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<p><font size = "5" face= "arial" color = "#184D79"><b>August 18-</b></font>
<p><font size = "5" face= "arial" color = "#184D79"><b>August 18-</b></font>
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             <blockquote><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"><li type = "circle"></blockquote>
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             <blockquote><li type = "circle">Glycerol Stock- 350 µL cells and 150 µL 50% glycerol<li type = "circle">Miniprep<li type = "circle">Test digestion<li type = "circle">Digestion, gel electrophoresis and purification<li type = "circle">Glycerol stock colonies 4 and 5 CheZ*<li type = "circle">Miniprep<li type = "circle">Test digestion and gel electrophoresis<li type = "circle">Ligation at 16 degrees Celsius overnight<li type = "circle">Incubation in 3 mL LB carb overnight at 37 degrees Celsius and 220 rpm<li type = "circle">Plate (507011) from Agar Stab</blockquote>
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Latest revision as of 02:14, 3 September 2011

biz solution

Week 6

August 15-

  • PCR of ptetR, RBS, LuxI, term
  • Gel electrophoresis and purification
  • Glycerol stock and miniprep
  • Incubation in 3 mL LB can overnight at 37 degrees Celsius and 220 rpm
  • Test digestion for 1 hour and 30 minutes at 37 degrees Celsius
  • Gel Electrophoresis
  • Test Digestion Observation: Lane 5 gave positive results for FGF-R: 2 bands observed at predicted bands size. Lane 4, 7, 9 were all negative bad results because EcoRI was used but only 1 band showed. This suggests that FGFR was not inserted. Lanes 10, 11, 12 were CheZ* and all matched negative control. Results are bad for CheZ*- will be regrown/miniprep/ligated.
  • Incubation in 3 mL LB chlora overnight at 37 degrees Celsius and 220 rpm
  • August 16-

  • Digestion; Gel Electrophoresis and Purification
  • Ligation (10 minutes at room temperature)
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • Re- incubating – taking 20 µL of yesterday’s incubations and placing them in 3 mL of LB and appropriate
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • August 17-

  • Glycerol stock ToxR and CheZ* colonies (2nd attempt)
  • Miniprep
  • no results on gel
  • Incubate in 3 mL LB carb overnight at 37 degrees Celsius and 220 rpm
  • Transformation (50 µL DH5α, 2 µL DNA, 37 degrees Celsius overnight, all carb)
  • Incubation in 3 mL of LB chlora overnight at 37 degrees Celsius and 220 rpm
  • August 18-

  • Glycerol Stock- 350 µL cells and 150 µL 50% glycerol
  • Miniprep
  • Test digestion
  • Digestion, gel electrophoresis and purification
  • Glycerol stock colonies 4 and 5 CheZ*
  • Miniprep
  • Test digestion and gel electrophoresis
  • Ligation at 16 degrees Celsius overnight
  • Incubation in 3 mL LB carb overnight at 37 degrees Celsius and 220 rpm
  • Plate (507011) from Agar Stab
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