Team:Warsaw/Calendar-Main/29 June 2009

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Making Llo fusion with secretion signal peptide

Kuba


Tasks:

  • Amplification of llo(hly) for fusion with the secretion signal peptide

Methods:

  • PCR mixture's composition:

    2,5ul pfu buffer (Fermentas)
    2,5ul MgSO4 (Fermentas)
    1,5ul primers, 1,5ul dNTPs (10 mM)
    0,5ul pfu turbo polymerase 
    1ul template DNA from Listeria, solution was topped up with H2O to 25ul

    One of the samples also contained 1 ul of DMSO

  • PCR programs:
  • hly

    4min 95°C 
    (30s 95°C, 40s 43°C, 1min30s 72°C)x3
    (30s 95°C, 40s 45°C, 1min30s 72°C)x28
    10min 72°C
    ~ 7°C

    random program

  • Electrophoretic separation on 1% agarose gel

Results:

  • Gel (from left)
  1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
  2. sample with DMSO
  3. sample without DMSO

  4. no products were observed



    Cloning of the mgtc promoter into the pKSII+ plasmid

    Kamil


    Tasks:

    • Purification of the PCR product
    • Plasmid assembly

    Methods:

    • The purification was carried out using Montage PCR Centrifugal Filter Device P36461 (Milipore) and according to the protocol supplied.

    • Plasmid digest mix was prepared as follows:
      2ul Tango buffer (Fermentas)
      1ul pKSII+ plasmid
      1ul XbaI enzyme
      1ul SmaI enzyme
      The solution was topped up with H2O to the final volume of 20 ul.
    • mgtc promoter digest mix was prepared as follows:
      2ul Tango buffer (Fermentas) 
      2ul purified gene
      1ul XbaI enzyme
      the solution was topped up with H2O to the final volume of 20 ul.
    • The digest was kept for 3h at 37°C, and then the enzyme was inactivated for 15min. at 80°C.
    • The ligation mix was prepared as follows:

      2ul digested plasmid 
      6ul mgtc digest product 
      2ul ligation buffer G (Fermentas)
      2ul T4 DNA ligase (Fermentas), 2ul 30% PEG
      2ul 10mM ATP
      The solution was topped up with H2O to the final volume of 20 ul. The ligation was carried out in 18°C overnight (~12h) and then inactivated for 10min. at 65°C.
    • A 200ul batch of chemocompetent bacteria was transformed with 500ul of ligation mix and incubated on petri dishes containing LB medium supplemented with ampicilin, X-gal and IPTG.


    Results:

    Due to a technological error the bacteria used were electrocompetent rather than chemocompetent. This resulted in there being not a single colony on the petri dishes.

    UPDATE: After another transformation and a second negative result a gel electrophoresis was performed revealing that the plasmid has not properly ligated.

    Electrophoresis results: Prom left:

    1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
    2. mgtc digest
    3. pKSII+ plasmid digest
    4. "ligation"





    5. April
      MTWTFSS
          [http://2009.igem.org/Team:Warsaw/Calendar-Main/1_April_2009 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_April_2009 2] [http://2009.igem.org/Team:Warsaw/Calendar-Main/3_April_2009 3] [http://2009.igem.org/Team:Warsaw/Calendar-Main/4_April_2009 4] [http://2009.igem.org/Team:Warsaw/Calendar-Main/5_April_2009 5]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/6_April_2009 6] [http://2009.igem.org/Team:Warsaw/Calendar-Main/7_April_2009 7] [http://2009.igem.org/Team:Warsaw/Calendar-Main/8_April_2009 8] [http://2009.igem.org/Team:Warsaw/Calendar-Main/9_April_2009 9] [http://2009.igem.org/Team:Warsaw/Calendar-Main/10_April_2009 10] [http://2009.igem.org/Team:Warsaw/Calendar-Main/11_April_2009 11] [http://2009.igem.org/Team:Warsaw/Calendar-Main/12_April_2009 12]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_April_2009 13] [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_April_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_April_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_April_2009 16] [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_April_2009 17] [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_April_2009 18] [http://2009.igem.org/Team:Warsaw/Calendar-Main/19_April_2009 19]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/20_April_2009 20] [http://2009.igem.org/Team:Warsaw/Calendar-Main/21_April_2009 21] [http://2009.igem.org/Team:Warsaw/Calendar-Main/22_April_2009 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_April_2009 23] [http://2009.igem.org/Team:Warsaw/Calendar-Main/24_April_2009 24] [http://2009.igem.org/Team:Warsaw/Calendar-Main/25_April_2009 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_April_2009 26]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/27_April_2009 27] [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_April_2009 28] [http://2009.igem.org/Team:Warsaw/Calendar-Main/29_April_2009 29] [http://2009.igem.org/Team:Warsaw/Calendar-Main/30_April_2009 30]
      May
      MTWTFSS
              [http://2009.igem.org/Team:Warsaw/Calendar-Main/1_May_2009 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_May_2009 2] [http://2009.igem.org/Team:Warsaw/Calendar-Main/3_May_2009 3]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/4_May_2009 4] [http://2009.igem.org/Team:Warsaw/Calendar-Main/5_May_2009 5] [http://2009.igem.org/Team:Warsaw/Calendar-Main/6_May_2009 6] [http://2009.igem.org/Team:Warsaw/Calendar-Main/7_May_2009 7] [http://2009.igem.org/Team:Warsaw/Calendar-Main/8_May_2009 8] [http://2009.igem.org/Team:Warsaw/Calendar-Main/9_May_2009 9] [http://2009.igem.org/Team:Warsaw/Calendar-Main/10_May_2009 10]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/11_May_2009 11] [http://2009.igem.org/Team:Warsaw/Calendar-Main/12_May_2009 12] [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_May_2009 13] [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_May_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_May_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_May_2009 16] [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_May_2009 17]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_May_2009 18] [http://2009.igem.org/Team:Warsaw/Calendar-Main/19_May_2009 19] [http://2009.igem.org/Team:Warsaw/Calendar-Main/20_May_2009 20] [http://2009.igem.org/Team:Warsaw/Calendar-Main/21_May_2009 21] [http://2009.igem.org/Team:Warsaw/Calendar-Main/22_May_2009 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_May_2009 23] [http://2009.igem.org/Team:Warsaw/Calendar-Main/24_May_2009 24]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/25_May_2009 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_May_2009 26] [http://2009.igem.org/Team:Warsaw/Calendar-Main/27_May_2009 27] [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_May_2009 28] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/29_May_2009&preload=Team:Warsaw/NotebookPreload&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/30_May_2009&preload=Team:Warsaw/NotebookPreload&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/31_May_2009&preload=Team:Warsaw/NotebookPreload&action=edit 31]
      June
      MTWTFSS
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/1_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/2_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/3_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/4_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/5_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/6_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/7_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 7]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/8_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/9_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/10_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/11_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/12_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/13_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/14_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 14]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/15_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_June_2009 16] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/17_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 17] [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_June_2009 18] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/19_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/20_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/21_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 21]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/22_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_June_2009 23] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/24_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/25_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_June_2009 26] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/27_June_2009&preload=Team:Warsaw/NotebookPreload&action=edit 27] [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_June_2009 28]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/29_June_2009 29] [http://2009.igem.org/Team:Warsaw/Calendar-Main/30_June_2009 30]
      July
      MTWTFSS
          [http://2009.igem.org/Team:Warsaw/Calendar-Main/1_July_2009 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_July_2009 2] [http://2009.igem.org/Team:Warsaw/Calendar-Main/3_July_2009 3] [http://2009.igem.org/Team:Warsaw/Calendar-Main/4_July_2009 4] [http://2009.igem.org/Team:Warsaw/Calendar-Main/5_July_2009 5]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/6_July_2009 6] [http://2009.igem.org/Team:Warsaw/Calendar-Main/7_July_2009 7] [http://2009.igem.org/Team:Warsaw/Calendar-Main/8_July_2009 8] [http://2009.igem.org/Team:Warsaw/Calendar-Main/9_July_2009 9] [http://2009.igem.org/Team:Warsaw/Calendar-Main/10_July_2009 10] [http://2009.igem.org/Team:Warsaw/Calendar-Main/11_July_2009 11] [http://2009.igem.org/Team:Warsaw/Calendar-Main/12_July_2009 12]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_July_2009 13] [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_July_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_July_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_July_2009 16] [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_July_2009 17] [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_July_2009 18] [http://2009.igem.org/Team:Warsaw/Calendar-Main/19_July_2009 19]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/20_July_2009 20] [http://2009.igem.org/Team:Warsaw/Calendar-Main/21_July_2009 21] [http://2009.igem.org/Team:Warsaw/Calendar-Main/22_July_2009 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_July_2009 23] [http://2009.igem.org/Team:Warsaw/Calendar-Main/24_July_2009 24] [http://2009.igem.org/Team:Warsaw/Calendar-Main/25_July_2009 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_July_2009 26]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/27_July_2009 27] [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_July_2009 28] [http://2009.igem.org/Team:Warsaw/Calendar-Main/29_July_2009 29] [http://2009.igem.org/Team:Warsaw/Calendar-Main/30_July_2009 30] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/31_July_2009&preload=Team:Warsaw/NotebookPreload&action=edit 31]
      August
      MTWTFSS
                [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/1_August_2009&preload=Team:Warsaw/NotebookPreload&action=edit 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_August_2009 2]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/3_August_2009 3] [http://2009.igem.org/Team:Warsaw/Calendar-Main/4_August_2009 4] [http://2009.igem.org/Team:Warsaw/Calendar-Main/5_August_2009 5] [http://2009.igem.org/Team:Warsaw/Calendar-Main/6_August_2009 6] [http://2009.igem.org/Team:Warsaw/Calendar-Main/7_August_2009 7] [http://2009.igem.org/Team:Warsaw/Calendar-Main/8_August_2009 8] [http://2009.igem.org/Team:Warsaw/Calendar-Main/9_August_2009 9]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/10_August_2009 10] [http://2009.igem.org/Team:Warsaw/Calendar-Main/11_August_2009 11] [http://2009.igem.org/Team:Warsaw/Calendar-Main/12_August_2009 12] [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_August_2009 13] [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_August_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_August_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_August_2009 16]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_August_2009 17] [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_August_2009 18] [http://2009.igem.org/Team:Warsaw/Calendar-Main/19_August_2009 19] [http://2009.igem.org/Team:Warsaw/Calendar-Main/20_August_2009 20] [http://2009.igem.org/Team:Warsaw/Calendar-Main/21_August_2009 21] [http://2009.igem.org/Team:Warsaw/Calendar-Main/22_August_2009 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_August_2009 23]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/24_August_2009 24] [http://2009.igem.org/Team:Warsaw/Calendar-Main/25_August_2009 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_August_2009 26] [http://2009.igem.org/Team:Warsaw/Calendar-Main/27_August_2009 27] [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_August_2009 28] [http://2009.igem.org/Team:Warsaw/Calendar-Main/29_August_2009 29] [http://2009.igem.org/Team:Warsaw/Calendar-Main/30_August_2009 30]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/31_August_2009 31]
      September
      MTWTFSS
        [http://2009.igem.org/Team:Warsaw/Calendar-Main/1_September_2009 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_September_2009 2] [http://2009.igem.org/Team:Warsaw/Calendar-Main/3_September_2009 3] [http://2009.igem.org/Team:Warsaw/Calendar-Main/4_September_2009 4] [http://2009.igem.org/Team:Warsaw/Calendar-Main/5_September_2009 5] [http://2009.igem.org/Team:Warsaw/Calendar-Main/6_September_2009 6]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/7_September_2009 7] [http://2009.igem.org/Team:Warsaw/Calendar-Main/8_September_2009 8] [http://2009.igem.org/Team:Warsaw/Calendar-Main/9_September_2009 9] [http://2009.igem.org/Team:Warsaw/Calendar-Main/10_September_2009 10] [http://2009.igem.org/Team:Warsaw/Calendar-Main/11_September_2009 11] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/12_September_2009&preload=Team:Warsaw/NotebookPreload&action=edit 12] [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_September_2009 13]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_September_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_September_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_September_2009 16] [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_September_2009 17] [http://2009.igem.org/Team:Warsaw/Calendar-Main/18_September_2009 18] [http://2009.igem.org/Team:Warsaw/Calendar-Main/19_September_2009 19] [http://2009.igem.org/Team:Warsaw/Calendar-Main/20_September_2009 20]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/21_September_2009 21] [http://2009.igem.org/Team:Warsaw/Calendar-Main/22_September_2009 22] [http://2009.igem.org/Team:Warsaw/Calendar-Main/23_September_2009 23] [http://2009.igem.org/Team:Warsaw/Calendar-Main/24_September_2009 24] [http://2009.igem.org/Team:Warsaw/Calendar-Main/25_September_2009 25] [http://2009.igem.org/Team:Warsaw/Calendar-Main/26_September_2009 26] [http://2009.igem.org/Team:Warsaw/Calendar-Main/27_September_2009 27]
      [http://2009.igem.org/Team:Warsaw/Calendar-Main/28_September_2009 28] [http://2009.igem.org/Team:Warsaw/Calendar-Main/29_September_2009 29] [http://2009.igem.org/Team:Warsaw/Calendar-Main/30_September_2009 30]
      October
      MTWTFSS
            [http://2009.igem.org/Team:Warsaw/Calendar-Main/1_October_2009 1] [http://2009.igem.org/Team:Warsaw/Calendar-Main/2_October_2009 2] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/3_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/4_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 4]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/5_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/6_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/7_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/8_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/9_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/10_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/11_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 11]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/12_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 12] [http://2009.igem.org/Team:Warsaw/Calendar-Main/13_October_2009 13] [http://2009.igem.org/Team:Warsaw/Calendar-Main/14_October_2009 14] [http://2009.igem.org/Team:Warsaw/Calendar-Main/15_October_2009 15] [http://2009.igem.org/Team:Warsaw/Calendar-Main/16_October_2009 16] [http://2009.igem.org/Team:Warsaw/Calendar-Main/17_October_2009 17] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/18_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 18]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/19_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/20_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/21_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/22_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/23_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/24_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/25_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 25]
      [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/26_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/27_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/28_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/29_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/30_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Warsaw/Calendar-Main/31_October_2009&preload=Team:Warsaw/NotebookPreload&action=edit 31]