Team:Goettingen/week6-2

From 2012.igem.org

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                            Keep it sync with the drop-down menu width.
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                            Keep it sync with the drop-down menu width.
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                            Use MenuTailor.css to customize. */
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/*--------------------------------------------------------------------------------------- Widths */
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/*
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/*
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Menu Bar 1
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Drop-Down Menu #2
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*/
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/*...............................................................................................*/
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/*
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Menu Bar 1
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Drop-Down Menu #2
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Side Menu #1
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*/
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                            Keep it sync with the drop-down menu width. */
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div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM1,
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/*...............................................................................................*/
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/*
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Menu Bar 1
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Drop-Down Menu #2
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Side Menu #2
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*/
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div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM2,
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                            Keep it sync with the drop-down menu width. */
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}
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div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM2,
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-
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-
/*...............................................................................................*/
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-
/*
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Menu Bar 1
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Drop-Down Menu #2
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Side Menu #3
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*/
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-
  left: 15.5em  !important; /* Places the side menu to the right of the drop-down menu.
+
-
                            Keep it sync with the drop-down menu width. */
+
-
}
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM3,
+
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-
  width: 17em; /* Side menu width. */
+
-
}
+
-
/*...............................................................................................*/
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-
 
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</style>
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</p><html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" dir="ltr" lang="en"><head>
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                <meta http-equiv="Content-Type" content="text/html; charset=UTF-8">
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                                <meta name="keywords" content="Team:Goettingen,Team:Goettingen,Team:Goettingen/Homing coli,Team:Goettingen/Human Practice/Flash coli,Team:Goettingen/Press,Team:Goettingen/Project/General information">
+
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                <link rel="shortcut icon" href="https://2012.igem.org/favicon.ico">
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Revision as of 09:14, 22 September 2012

Deutsch  / English 

Language: English

#2 Speed Improvement - 6th week

Back to overview

V06_04


Chemical retransformation of the flhDC-promoter constructs into E. coli (DH10B)
  • Experiment:
    For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:

    20E - #2
    20E - #3
    18M - #2
    18M - #3
    18C - #1
    18C - #2

  • Observations & Results:
    The retransformation was successful since all plates showed numeours colonies except the negative control.


V06_05


V06_05_1 Preparative double digestion of flhDC and plasmids containing further promoters
  • Experiment:
    In order to clone flhDC behind further promoters, the purified PCR product was digested with XbaI and PstI, whereas the BioBrick plasmids were cut with SpeI and PstI. The digestion was performed as described in the protocoll. This time the follwing promoters were used:

    J23113 - 20G - x21
    J23109 - 2G - x106
    J23114 - 20I - x256
    J23106 - 18O - x1185
    J23104 - 18K - x 1831


V06_05_2 Insertion of flhDC into the BioBrick plamids
  • Experiment:
    The ligation of the digested flhDC with the BioBrick plasmids was conducted as described in the protocol.


V06_06


Chemical transformation of the flhDC-promoter constructs into E. coli (DH10B)
  • Experiment:
    For the chemical transformation the standard protocol was followed.
  • Observations & Results:
    The transformation was successful since all plates showed numeours colonies except the negative control. However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected.


V06_07


Preparation of over night cultures
  • Experiment:
    Over night cultures were prepapred of three colonies of each constructs in order to isolate the plasmids.


V06_08


V06_08_1 Miniprep of the new flhDC-promoter constructs
  • Experiment:
    Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

    V06_08_2 Test digestion of the new flhDC-promoter constructs
  • Experiment:
    In order to prove correct insertion of flhDC a test digestion was performed using SpeI and XbaI according to the protocol.
  • Observations & Results:
    For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.


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Important pages:
Home; Team; Official Team Profile; Project; Parts submitted to the Registry; Modeling; Notebook; Saftey; Attributions

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