Team:Warsaw/Calendar-Main/18 August 2009

From 2009.igem.org

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<h3><div style="text-align: center;">Cloning switch 1 regulatory parts [
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012">K177012</a>-PcI.RBS.LacI,
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177033">K177033</a>-PcI.RBS.LacI.PcI.RBS.RFP.terminator,
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177011">K177011</a>-PLacI.RBS.cI.terminator,
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<a href="http://partsregistry.org/Part:BBa_K177038">K177038</a>-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator
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]
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into two compatible low copy number plasmids of different antibiotic resistance</div></h3>
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<h4>Ania</h4>
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<p>Tasks:</p>
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Revision as of 00:10, 10 September 2009


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Assembly of endosomal detection operon

Marcin


Task 1:

Methods:

  • All samples were inactivated via heating in 80 °C for 20 minutes
  • All gel-outs were performed using the EurX gel-out kit according to the manual

Results

  • All gel-outs (except C0040 with RBS) were successful. One isolation failed because of very low DNA concentration

Task 2:

  • Digest pKSII plasmid with cro CDS

Methods:

  • Reaction mixture composition:
  •  
    45 μl purified plasmid DNA product
    1 μl XbaI (Fermentas)
    0.5 μl SpeI (Fermentas)
    5 μl Buffer Tango (Fermentas)

    Task 3:

    Methods:

    • Reaction mixture composition:
    • 10 μl p53 CDS
      7 μl vector
      2.3 μl Buffer Tango (Fermentas)
      3 μl dNTPs mixture (EurX)
      1 μl T4 ligase (Fermentas)
      
    • Ligation was performed for about 9 hours and it was subsequently thermally inactivated


    Making of plac-RBS-llo part

    Jarek


    Tasks:

    • Preparation of 20 liquid cultures, from colonies that grew from ligation, in 3 ml of LB+Amp nad incubating them in 37C degree

    Cloning switch 1 regulatory parts [ K177012-PcI.RBS.LacI, K177033-PcI.RBS.LacI.PcI.RBS.RFP.terminator, K177011-PLacI.RBS.cI.terminator, K177038-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator ] into two compatible low copy number plasmids of different antibiotic resistance

    Ania


    Tasks:


    April
    MTWTFSS
        1 2 3 4 5
    6 7 8 9 10 11 12
    13 14 15 16 17 18 19
    20 21 22 23 24 25 26
    27 28 29 30
    May
    MTWTFSS
            1 2 3
    4 5 6 7 8 9 10
    11 12 13 14 15 16 17
    18 19 20 21 22 23 24
    25 26 27 28 29 30 31
    June
    MTWTFSS
    1 2 3 4 5 6 7
    8 9 10 11 12 13 14
    15 16 17 18 19 20 21
    22 23 24 25 26 27 28
    29 30
    July
    MTWTFSS
        1 2 3 4 5
    6 7 8 9 10 11 12
    13 14 15 16 17 18 19
    20 21 22 23 24 25 26
    27 28 29 30 31
    August
    MTWTFSS
              1 2
    3 4 5 6 7 8 9
    10 11 12 13 14 15 16
    17 18 19 20 21 22 23
    24 25 26 27 28 29 30
    31
    September
    MTWTFSS
      1 2 3 4 5 6
    7 8 9 10 11 12 13
    14 15 16 17 18 19 20
    21 22 23 24 25 26 27
    28 29 30
    October
    MTWTFSS
          1 2 3 4
    5 6 7 8 9 10 11
    12 13 14 15 16 17 18
    19 20 21 22 23 24 25
    26 27 28 29 30 31