Team:Goettingen/Project/Materials

From 2012.igem.org

(Difference between revisions)
 
(58 intermediate revisions not shown)
Line 1: Line 1:
-
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" dir="ltr" lang="en"><head>
+
{{GoettingenHeader|deu=Team:Goettingen/Project/Materials|eng=Team:Goettingen/Project/Materials}}
-
                <meta http-equiv="Content-Type" content="text/html; charset=UTF-8">
+
{{GoettingenBlue}}
-
                                <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">
+
-
                <link rel="shortcut icon" href="https://2012.igem.org/favicon.ico">
+
-
                <link rel="search" type="application/opensearchdescription+xml" href="https://2012.igem.org/wiki/opensearch_desc.php" title="2012.igem.org (English)">
+
-
                <link title="Creative Commons" type="application/rdf+xml" href="https://2012.igem.org/wiki/index.php?title=Team:Goettingen&amp;action=creativecommons" rel="meta">
+
-
<link rel="alternate" type="application/rss+xml" title="2012.igem.org RSS Feed" href="https://2012.igem.org/wiki/index.php?title=Special:Recentchanges&amp;feed=rss">
+
-
<link rel="alternate" type="application/atom+xml" title="2012.igem.org Atom Feed" href="https://2012.igem.org/wiki/index.php?title=Special:Recentchanges&amp;feed=atom">
+
 +
== Recipes ==
 +
=== Media ===
 +
Recipes for liquid and solid media for bacterial cultivation. Stir until dissolved; sterilize by autoclaving 121 °C. Do not forget a magnetic stirrer.
-
<body class="mediawiki ns-0 ltr page-Team_Goettingen">
+
==== LB-broth ====
 +
* 1 % (w/v) bacto tryptone
 +
* 0.5 % (w/v) bacto yeast extract
 +
* 1 % (w/v) NaCl
 +
* if required: ad 1 % (w/v) agar
 +
==== M9 Swimming Agar ====
 +
* 1.25 % (v/v) glycerol
 +
* 20 % (v/v) 5x M9 salt stock solution
 +
* 0.1 % (v/v) of CaCl<sub>2</sub> x 2 H<sub>2</sub>O stock solution (20 mg/mL) 
 +
* 0.1 % (v/v) MgSO<sub>4</sub> stock solution (0.12 g/mL) 
 +
* 0.3 % (w/v) of agar
 +
==== Tryptone swimming agar ====
 +
* 1 % (w/v) bacto tryptone
 +
* 0.5 % (w/v) NaCl
 +
* 0.3 % (w/v) agar
 +
=== Antibiotics ===
 +
* Stock solutions and concentrations of antibiotics.
-
                        <!-- start content -->
+
{| class="goetable"
 +
|-
 +
! Reagent !!Abbreviation !! Stock Conc. [mg/mL] !! Working Conc. [µg/mL]!! Dilution !! Solvent         !!style="width:400px"|Notes
 +
|-
 +
|Ampicillin ||Amp ||style="text-align:right"| 100 ||style="text-align:right"| 100 ||style="text-align:right"| 1,000x ||ddH<sub>2</sub>O                 ||Culture plates with Amp can be stored at 4 °C for about 2 weeks. Stock solutions can be stored at 4 °C for 2 weeks but can last as long as 4-6 months when stored at -20 °C.
 +
|-
 +
|Chloramphenicol ||Cm ||style="text-align:right"| 20  ||style="text-align:right"| 20 ||style="text-align:right"| 1,000x ||EtOH                         ||
 +
|- 
 +
|Kanamycin ||Kan ||style="text-align:right"| 50  ||style="text-align:right"| 50 ||style="text-align:right"| 1,000x ||ddH2O                 ||Plates may only last a few weeks at 4 °C.
 +
|}
 +
=== Supplements ===
 +
* <u>Aminoacid mix</u> (containing all aminoacids except tryptophane): stocksolution 0.2 % (w/v), varying amounts used in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] and notebook of group 1
 +
* <u>Caffeine</u>: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]]
 +
* <u>D-Aspartate</u>: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]]
 +
* <u>2-Ethyl-1-Hexanole</u>: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] 
 +
* <u>Geraniol</u>: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] 
 +
* <u>L-Aspartate-4-Benzylester</u>: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] 
 +
* <u>L-aspartate</u>: stocksolution 10 mM, varying amounts used in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] and notebook of group 1
 +
* <u>Leucin</u>: stocksolution 30.49 mM, add to M9-agar: 4 mL (final concentration 0.305 µM) 
 +
* <u>Methionin</u>: stocksolution 26.8 mM, add to M9-agar: 4 mL (final concentration 0.268 µM) 
 +
* <u>Sodium Cyclamate</u>: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] 
 +
* <u>Tryptone</u>: stocksolution 0.5 % (w/v), varying amounts used in swimming assays, view [[Team:Goettingen/Project/Methods|methods]] and notebook of group 1 
 +
* <u>Vanillin</u>: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view [[Team:Goettingen/Project/Methods|methods]]
-
<style>
+
=== Buffer and Solution ===
-
h1.firstHeading { display: none; }
+
Recipes for stocks of buffers and solutions.
-
p {text-align: justify;}
+
==== 5x M9 salts stock solution ====
 +
{| class="goetable"
 +
|-
 +
!Amount !! Substance
 +
|-
 +
|64 g || Na<sub>2</sub>HPO<sub>4</sub> &times; 7 H<sub>2</sub>O
 +
|-
 +
|15 g || KH<sub>2</sub>PO<sub>4</sub>
 +
|-
 +
|2.5 g || NaCl
 +
|-
 +
|5.0 g || NH<sub>4</sub>Cl
 +
|-
 +
|ad 1000 ml || ddH<sub>2</sub>O
 +
|-
 +
|}
 +
* Stir until dissolved
-
a:link { color: #004080; text-decoration: none}
+
==== CaCl<sub>2</sub> Buffer for competent cells ====
-
a:visited { color:##003090; text-decoration: none}
+
{| class="goetable"
-
a:hover { color:#f29400; text-decoration: none}
+
|-
-
a:active { color:#f29400; text-decoration: none}
+
!Amount !! Substance !! Notes
 +
|-
 +
| 7.351 g|| CaCl<sub>2</sub> &times; 7 H<sub>2</sub>O || Alternative 0.1 M CaCl<sub>2</sub>
 +
|-
 +
|15%  || Glycerol ||
 +
|-
 +
|ad 500 ml || ddH<sub>2</sub>O ||
 +
|-
 +
|}
 +
* Stir until dissolved; sterilize by sterile filtration.
-
#bodyContent { padding: 10px auto; width: 910px; margin: auto; clear: none; }
+
==== 50x TAE for agarose gels ====
 +
{| class="goetable"
 +
|-
 +
! Amount !! Substance
 +
|-
 +
|242 g || Tris base
 +
|-
 +
|57.1 mL || Glacial acetic acid
 +
|-
 +
|18.6 g || EDTA
 +
|-
 +
|ad 1000 mL || ddH<sub>2</sub>O
 +
|-
 +
|}
 +
* Stir until dissolved. Working concentration is 1x (40 mL of 50x stock ad 2 L ddH<sub>2</sub>O).
-
table#team_members { text-align: justify; border: 0; }
+
== <i>E. coli</i> ==
-
table#team_members h2, table#team_members h3 { clear: both; }
+
*<i>E. coli</i> strain list - List of the working <i>E. coli</i> strains including genotypes, genetic markers, and alleles.
-
 
+
=== Non-mutated strains ===
-
 
+
These strains may not contain any plasmids or resistances, because they should be used for transformation or other experiments.
-
/*-----------------------------------------------------------------------------------------------*/
+
-
div.MenuBar ul li ul.DropDownMenu {
+
-
  display: none; /* Hides all drop-down menus. */
+
-
 
+
-
}
+
-
/*
+
-
li:hover works in IE7 and FF2.
+
-
a:hover works in IE6 and FF2.
+
-
a:hover breaks li:hover in FF2.
+
-
*/
+
-
div.MenuBar ul li:hover ul.DropDownMenu li ul.SideMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a ul.SideMenu {
+
-
  display: none; /* Hides all side menus. */
+
-
}
+
-
/*------------------------------------------------------------------------------------- Menu Bar */
+
-
div.MenuBar {
+
-
  font: Verdana;
+
-
  height: 30px;
+
-
  width: 910px;
+
-
  /*width: 100%*/
+
-
  margin: 0;
+
-
  border-top: 0;
+
-
  border-right: 0;
+
-
  border-left: 0;
+
-
  padding: 0;
+
-
  background: #00446b;
+
-
 
+
-
}
+
-
div.MenuBar ul {
+
-
  font: Verdana;
+
-
  text-align: center;
+
-
  list-style-type: none;
+
-
  margin: 0 auto;
+
-
  border: 0;
+
-
  padding: 0;
+
-
  background: #00446b;
+
-
}
+
-
div.MenuBar ul li {
+
-
  font: Verdana;
+
-
  display: block;
+
-
  padding: 0;
+
-
  margin: 0;
+
-
  font-size: 1.3em;
+
-
  float: left;
+
-
  background: #00446b;
+
-
  text-align: center;
+
-
  width: 107px;
+
-
  position: relative; /* Sets the positioning context for each drop-down menu. */
+
-
}
+
-
 
+
-
div.MenuBar ul li a {
+
-
  font: Verdana;
+
-
  display: block;
+
-
  background: #00446b;
+
-
  height: 22px; /* Keep height + padding-top + padding-bottom sync with the menu bar height. */
+
-
  color: white;
+
-
  padding-top: 4px;
+
-
  padding-bottom: 4px;
+
-
  padding-left: 1em; /* Sets the left space between top-level items. */
+
-
  padding-right: 1em; /* Sets the right space between top-level items. */
+
-
  text-decoration: none;
+
-
}
+
-
 
+
-
/*------------------------------------------------------------------------------ Drop-Down Menus */
+
-
div.MenuBar ul li:hover ul.DropDownMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu {
+
-
  display: block;
+
-
  width: 10em; /* Drop-down menu width.
+
-
                  Use MenuTailor.css to customize. */
+
-
  height: 1em;
+
-
  padding: 1px; /* Sets the drop-down menu "effective border" width. */
+
-
  position: absolute;
+
-
  top: 28px; /* Places the drop-down menu under the menu bar.
+
-
                Keep it sync with the menu bar height. */
+
-
  left: 0; /* Aligns the drop-down menu to its top-level item. */
+
-
  background-color: #A3C0E0; /* Selected item. */
+
-
  color: #FFFFFF;
+
-
 
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a {
+
-
  width: 10em; /* Keep it sync with the drop-down menu width.
+
-
                            Use MenuTailor.css to customize. */
+
-
  height: 1em;
+
-
  padding-left: 0;
+
-
  padding-right: 0;
+
-
  background-color: #A3C0E0; /* Selected item. */
+
-
  color: #FFFFFF;
+
-
}
+
-
ul.DropDownMenu li a span {
+
-
  display: block;
+
-
  padding-left: 0.75em; /* Sets the left space of each drop-down menu item. */
+
-
  padding-right: 0.25em; /* Sets the right space of each drop-down menu item. */
+
-
  text-align: right; /* Aligns the >> symbol to the right. */
+
-
}
+
-
ul.DropDownMenu li a span span {
+
-
  float: left; /* Aligns the text (back) to the left. */
+
-
  font: 12px Verdana; /* Required for IE55. */
+
-
  height: 20px;
+
-
  color: #FFFFFF;
+
-
}
+
-
/*----------------------------------------------------------------------------------- Side Menus */
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu {
+
-
  display: block;
+
-
  width: 11em; /* Side menu width.
+
-
                  Use MenuTailor.css to customize. */
+
-
  padding: 1px; /* Sets the side menu "effective border" width. */
+
-
  position: absolute;
+
-
  top: -1px; /* Aligns the side menu to its drop-down menu item.
+
-
                Keep it sync with the side menu "effective border" width. */
+
-
  left: 13em; /* Places the side menu to the right of the drop-down menu.
+
-
                            Keep it sync with the drop-down menu width.
+
-
                            Use MenuTailor.css to customize. */
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu li a {
+
-
  width: 11em; /* Keep it sync with the side menu width.
+
-
                            Use MenuTailor.css to customize. */
+
-
  font: 12px Verdana; /* Required for IE55. */
+
-
  left: 13em; /* Places the side menu to the right of the drop-down menu.
+
-
                            Keep it sync with the drop-down menu width.
+
-
                            Use MenuTailor.css to customize. */
+
-
}
+
-
div.MenuBar ul li ul.DropDownMenu li ul.SideMenu li a span {
+
-
  padding-left: 1.5em; /* Sets the left space of each side menu item. */
+
-
  padding-right: 0.5em; /* Sets the right space of each side menu item. */
+
-
  text-align: left;
+
-
  font: 12px Verdana; /* Required for IE55. */
+
-
  left: 13em; /* Places the side menu to the right of the drop-down menu.
+
-
                            Keep it sync with the drop-down menu width.
+
-
                            Use MenuTailor.css to customize. */
+
-
}
+
-
/*----------------------------------------------------------------------------- Browser Specific */
+
-
* html div.MenuBar ul li a {
+
-
  float: left; /* Required for IE55 and IE6.
+
-
                            Breaks O9.
+
-
                            Hidden (* html) from non-IE browsers. */
+
-
}
+
-
* html ul.DropDownMenu li a:hover {
+
-
  cursor: hand; /* Required for IE55.
+
-
                  Hidden (* html) from non-IE browsers. */
+
-
}
+
-
ul.DropDownMenu li a:hover {
+
-
  cursor: pointer; /* Required for IE6 and IE7.
+
-
                      Hidding it (* html) from non-IE browsers breaks IE7!
+
-
}
+
-
* html div.MenuBar a:hover {
+
-
  text-decoration: none; /* Required for IE55 and IE6.
+
-
                            Hidden (* html) from non-IE browsers. */
+
-
}
+
-
* html div.MenuBar ul li table,
+
-
* html div.MenuBar ul li table td {
+
-
  border: 0; /* Required for IE55 and IE6.
+
-
                Hidden (* html) from non-IE browsers. */
+
-
}
+
-
/*------------------------------------------------------------------------------- Default Colors */
+
-
div.MenuBar {
+
-
  background-color: Menu;
+
-
  border-bottom: 1px solid ButtonShadow;
+
-
}
+
-
div.MenuBar a {
+
-
  background-color: Menu; /* Top-level unselected items. */
+
-
  color: MenuText;
+
-
}
+
-
div.MenuBar ul li:hover a,
+
-
div.MenuBar ul li a:hover {
+
-
  color: #A3C0E0;
+
-
  background-color: Highlight; /* Top-level selected item. */
+
-
  color: HighlightText;
+
-
}
+
-
/*...............................................................................................*/
+
-
div.MenuBar ul li:hover ul.DropDownMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu {
+
-
  background-color: ButtonShadow; /* Sets the drop-down menu "effective border" color. */
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a {
+
-
  background-color: Menu;  Drop-down menu unselected items.
+
-
                            Sets the drop-down menu "effective background" color. */
+
-
  color: MenuText;
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover {
+
-
  background-color: Highlight; /* Drop-down menu selected item. */
+
-
  color: HighlightText;
+
-
}
+
-
/*...............................................................................................*/
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu {
+
-
  background-color: ButtonShadow; /* Sets the side menu "effective border" color. */
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu li a {
+
-
  background-color: Menu; /* Side menu unselected items.
+
-
                            Sets the side menu "effective background" color. */
+
-
  color: MenuText;
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a:hover,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu li a:hover {
+
-
  background-color: Highlight; /* Side menu selected item. */
+
-
  color: HighlightText;
+
-
}
+
-
/*-----------------------------------------------------------------------------------------------*/
+
-
 
+
-
/*-------------------------------------------------------------------------------------- General */
+
-
body {
+
-
  background: white;
+
-
  color: black;
+
-
  margin: 0;
+
-
  border: 0;
+
-
  padding: 0;
+
-
}
+
-
 
+
-
 
+
-
div.MenuBar {
+
-
  font: 13px arial, helvetica, sans-serif;
+
-
}
+
-
div.MenuBar ul {
+
-
  font: 13px arial, helvetica, sans-serif; /* Required for IE55. */
+
-
}
+
-
/*--------------------------------------------------------------------------------------- Colors */
+
-
div.MenuBar {
+
-
  background-color: #0064C8; /* Selected item. */
+
-
  color: #FFFFFF;
+
-
  border-bottom: 1px solid ButtonShadow;
+
-
}
+
-
div.MenuBar a,
+
-
div.MenuBar ul li:hover ul.DropDownMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a,
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu li a {
+
-
  background-color: #00446b; /* Selected item. */
+
-
  color: #FFFFFF;
+
-
}
+
-
div.MenuBar ul li:hover a,
+
-
div.MenuBar ul li a:hover,
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover a,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover,
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a:hover,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu li a:hover {
+
-
  background-color: #A3C0E0; /* Selected item. */
+
-
  color: #FFFFFF;
+
-
}
+
-
div.MenuBar ul li:hover ul.DropDownMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu,
+
-
div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu,
+
-
div.MenuBar ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu {
+
-
  background-color: ButtonShadow; /* Sets the drop-down and side menus "effective border" color. */
+
-
}
+
-
/*--------------------------------------------------------------------------------------- Widths */
+
-
/*
+
-
 
+
-
/*
+
-
Menu Bar 1
+
-
Drop-Down Menu #2
+
-
*/
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu#MB1-DDM4,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu#MB1-DDM4,
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu#MB1-DDM4 li a,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu#MB1-DDM4 li a {
+
-
  width: 15.5em; /* Drop-down menu width. */
+
-
}
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu#MB1-DDM5,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu#MB1-DDM5,
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu#MB1-DDM5 li a,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu#MB1-DDM5 li a {
+
-
  width: 14em; /* Drop-down menu width. */
+
-
}
+
-
 
+
-
/*...............................................................................................*/
+
-
/*
+
-
Menu Bar 1
+
-
Drop-Down Menu #2
+
-
Side Menu #1
+
-
*/
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM1,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM1 {
+
-
  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-SM1,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM1,
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM1 li a,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM1 li a {
+
-
  width: 15em; /* Side menu width. */
+
-
}
+
-
/*...............................................................................................*/
+
-
/*
+
-
Menu Bar 1
+
-
Drop-Down Menu #2
+
-
Side Menu #2
+
-
*/
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM2,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM2 {
+
-
  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-SM2,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM2,
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM2 li a,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM2 li a {
+
-
  width: 7em; /* Side menu width. */
+
-
}
+
-
/*...............................................................................................*/
+
-
/*
+
-
Menu Bar 1
+
-
Drop-Down Menu #2
+
-
Side Menu #3
+
-
*/
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM3,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM3 {
+
-
  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,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM3,
+
-
div.MenuBar#navi ul li:hover ul.DropDownMenu li:hover ul.SideMenu#MB1-DDM2-SM3 li a,
+
-
div.MenuBar#navi ul li a:hover ul.DropDownMenu li a:hover ul.SideMenu#MB1-DDM2-SM3 li a {
+
-
  width: 17em; /* Side menu width. */
+
-
}
+
-
/*...............................................................................................*/
+
-
 
+
-
</style>
+
-
 
+
-
 
+
-
 
+
-
</p><html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" dir="ltr" lang="en"><head>
+
-
                <meta http-equiv="Content-Type" content="text/html; charset=UTF-8">
+
-
                                <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">
+
-
                <link rel="shortcut icon" href="https://2012.igem.org/favicon.ico">
+
-
                <link rel="search" type="application/opensearchdescription+xml" href="https://2012.igem.org/wiki/opensearch_desc.php" title="2012.igem.org (English)">
+
-
                <link title="Creative Commons" type="application/rdf+xml" href="https://2012.igem.org/wiki/index.php?title=Team:Goettingen&amp;action=creativecommons" rel="meta">
+
-
<link rel="alternate" type="application/rss+xml" title="2012.igem.org RSS Feed" href="https://2012.igem.org/wiki/index.php?title=Special:Recentchanges&amp;feed=rss">
+
-
<link rel="alternate" type="application/atom+xml" title="2012.igem.org Atom Feed" href="https://2012.igem.org/wiki/index.php?title=Special:Recentchanges&amp;feed=atom">
+
-
 
+
-
 
+
-
<body class="mediawiki ns-0 ltr page-Team_Goettingen">
+
-
 
+
-
 
+
-
 
+
-
 
+
-
                        <!-- start content -->
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<div id="header"><img src="http://www.patrickreinke.de/igem/headpicture_2.jpg" alt="Team Goettingen">
+
-
<br><div class="MenuBar" id="navi">
+
-
        <ul>
+
-
                <li>
+
-
                      <a href="https://2012.igem.org/Team:Goettingen" style="color: white;">Home<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                        <ul class="DropDownMenu" id="MB1-DDM2-SM3">
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/News"><span><span>News</span></span></a></li>
+
-
                        <!--<li><a href="https://2012.igem.org/Team:Goettingen/News"><span><span><div style="text-indent:20px;">&#8901; Advisors</div></span></span></a></li> -->
+
-
                        </ul>
+
-
                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Team" style="color: white;">Team<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                        <ul class="DropDownMenu" id="MB1-DDM5">
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Team"><span><span>Members</span></span></a></li>
+
-
                        <!--<li><a href="https://2012.igem.org/Team:Goettingen/Team#advisor"><span><span><div style="text-indent:20px;">&#8901; Advisors</div></span></span></a></li> -->
+
-
                        <!--<li><a href="https://2012.igem.org/Team:Goettingen/Team#students"><span><span><div style="text-indent:20px;">&#8901; Students</div></span></span></a></li> -->
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/Team#responsibitlities"><span><span><div style="text-indent:20px;">&#8901; Responsibilities</div></span></span></a></li> -->
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Focus_Groups"><span><span>Focus Groups</span></span></a></li>
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/groups#group1"><span><span><div style="text-indent:20px;">&#8901; #1 Selection / Swimming</div></span></span></a></li>  -->
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/groups#group2"><span><span><div style="text-indent:20px;">&#8901; #2 Speed Improvement</div></span></span></a></li>  -->
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/groups#group3"><span><span><div style="text-indent:20px;">&#8901; #3 Chemoreceptor</div></span></span></a></li>    -->
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/groups#library"><span><span><div style="text-indent:20px;">&#8901; Library</div></span></span></a></li>            -->
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Seminars"><span><span>Seminars</span></span></a></li>
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/seminars#announce"><span><span><div style="text-indent:20px;">&#8901; Announcements</div></span></span></a></li>    -->
+
-
                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/seminars#presentations"><span><span><div style="text-indent:20px;">&#8901; Past Presentations</div></span></span></a></li> -->
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Work_Impressions"><span><span>Work Impressions</span></span></a></li>
+
-
                        </ul>
+
-
                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Project" style="color: white;">Project<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                        <ul class="DropDownMenu" id="MB1-DDM2-SM3">
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project"><span><span>Our Project</span></span></a></li>
+
-
                        <!--<li></li><li><a href="https://2012.igem.org/Team:Goettingen/Chemotaxis"><span><span><div style="text-indent:20px;">&#8901; Chemotaxis</div></span></span></a></li>  -->
+
-
                        <!--<li></li><li><a href="https://2012.igem.org/Team:Goettingen/Poster"><span><span><div style="text-indent:20px;">&#8901; Poster</div></span></span></a></li>    -->
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Materials"><span><span>Materials</span></span></a>
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Methods"><span><span>Methods</span></span></a>
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Computational_Data"><span><span>Computational Data</span></span></a>
+
-
                        <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Maps"><span><span><div style="text-indent:20px;">&#8901; Maps</div></span></span></a></li>
+
-
                        </li><li><a href="https://2012.igem.org/Team:Goettingen/Sequences"><span><span><div style="text-indent:20px;">&#8901; Sequences</div></span></span></a></li> -->
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Bioinformatical_Tool"><span><span>Bioinformatical Tools</span></span></a>
+
-
                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Lab_Work_Flow"><span><span>Lab Work Flow</span></span></a>
+
-
                        <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Maps"><span><span><div style="text-indent:20px;">&#8901; Golden Lab Rules</div></span></span></a></li>
+
-
                        </li><li><a href="https://2012.igem.org/Team:Goettingen/Sequences"><span><span><div style="text-indent:20px;">&#8901; Daily Check List</div></span></span></a></li> -->
+
-
                        </ul>
+
-
                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
 
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Notebook" style="color: white;">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                <ul class="DropDownMenu" id="MB1-DDM2">
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook"><span><span>Overview</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook/Summary"><span><span>Summary</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook/Discussion"><span><span>Discussion</span></span></a></li>
+
-
                                </ul>
+
-
                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/iGEM" style="color: white;">iGEM<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                <ul class="DropDownMenu" id="MB1-DDM6">
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM"><span><span>What is iGEM?</span></span></a></li>
+
-
                                <!--<li><a href="https://2012.igem.org/Team:Goettingen/BioBrick_System"><span><span>iGEM BioBrick System</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Part_Registry"><span><span>iGEM Part Registry</span></span></a></li>  -->
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Parts"><span><span>Parts Submitted</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Attributions"><span><span>Attributions</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Saftey"><span><span>Safety</span></span></a></li>>
+
-
                                </ul>
+
-
                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Goettingen" style="color: white;">G&ouml;ttingen<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                <ul class="DropDownMenu" id="MB1-DDM5">
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen"><span><span>City</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/University"><span><span>Georg-August-University</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/MPI"><span><span>Max-Planck-Institute</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/S1-Demo_Lab"><span><span>S1-Demo Lab</span></span></a></li>
+
-
 
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                                </li>
+
-
 
+
-
                                </ul>
+
-
                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
                <li style="width: 160px">
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Human_Practice" style="color: white;">Human Practice<!--[if gt IE 6]><!--></a><!--<![endif]-->
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                <ul class="DropDownMenu" id="MB1-DDM5">
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice"><span><span>Why Human Practice?</span></span></a></li>
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Public_and_Media"><span><span>Public and Media</span></span></a></li>
+
-
                              <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Newspaper"><span><span><div style="text-indent:20px;">&#8901; Newspaper</div></span></span></a></li>
+
-
                              </li><li><a href="https://2012.igem.org/Team:Goettingen/Conference"><span><span><div style="text-indent:20px;">&#8901; Conference</div></span></span></a></li>
+
-
                              </li><li><a href="https://2012.igem.org/Team:Goettingen/Synthetic_Biology_Day"><span><span><div style="text-indent:20px;">&#8901; Synthetic Biology Day</div></span></span></a></li> -->
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Survey"><span><span>Survey</span></span></a></li>
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Synthetic_Biology_Day"><span><span>Synthetic Biology Day</span></span></a></li>
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Panel_Discussion"><span><span>Panel Discussion</span></span></a></li>-->
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Flash_coli"><span><span>Flash Coli</span></span></a></li>
+
-
                              </ul>
+
-
                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
+
-
                </li>
+
-
 
+
-
                <li>
+
-
                        <a href="https://2012.igem.org/Team:Goettingen/Sponsoring" style="color: white;">Sponsoring</a>
+
-
                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
+
-
                                <ul class="DropDownMenu" id="MB1-DDM2-SM3">
+
-
                              <li><a href="https://2012.igem.org/Team:Goettingen/Sponsoring"><span><span>Interested in...?</span></span></a></li>
+
-
                                <!-- <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Sponsors"><span><span>Sponsors</span></span></a></li>
+
-
                                <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Supporters"><span><span>Supporters</span></span></a></li>    -->
+
-
                </li>
+
-
        </ul>
+
-
 
+
-
</div>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<table>
+
-
 
+
-
                                <!-- Text body -->
+
-
<tbody><tr valign="top" align="left">
+
-
 
+
-
<td style="padding: 0pt 20px 0pt 0pt;" width="650px">
+
-
<font face="Verdana" size="-1">
+
<br>
<br>
-
Language: <img height="20", src="http://www.patrickreinke.de/igem/eng.jpg"> English <br>
+
==== DH10B ====
 +
===== Genotype =====
 +
*<i>F- endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74 Φ80lacZΔM15 araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) λ- </i>
 +
===== Genetical Characteristics =====
 +
* Antibiotic resistance: none
 +
===== Reference =====
 +
* <html><a href="http://products.invitrogen.com/ivgn/product/18297010">Invitrogen</a></html>
<br>
<br>
-
 
+
==== DH5α ====
-
<table id="toc" class="toc"><tbody><tr><td><div id="toctitle"><h2>Contents</h2> <span class="toctoggle"><a href="javascript:toggleToc()" class="internal" id="togglelink"></a></span></div>
+
===== Genotype =====
-
<ul>
+
*<i>F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ–</i>
-
<li class="toclevel-1"><a href="#Recipes"><span class="tocnumber">1</span> <span class="toctext">Recipes</span></a></li>
+
===== Genetical Characteristics =====
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Media"><span class="tocnumber"></span> <span class="toctext">Media</span></a></li>
+
* Antibiotic resistance: none
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Antibiotics"><span class="tocnumber"></span> <span class="toctext">Antibiotics</span></a></li>
+
===== Reference =====
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Supplements"><span class="tocnumber"></span> <span class="toctext">Supplements</span></a></li>
+
* <html><a href="http://www.invitrogen.com/1/3/dh5a-competent-cells">Invitrogen</a></html>
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Buffer_and_Solutions"><span class="tocnumber"></span> <span class="toctext">Buffer and Solutions</span></a></li>
+
-
<li class="toclevel-1"><a href="#E._coli"><span class="tocnumber">2</span> <span class="toctext"><i>E. coli</i></span></a></li>
+
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Strains"><span class="tocnumber"></span> <span class="toctext">Strains</span></a></li>
+
-
<li class="toclevel-1"><div style="text-indent:10px;"><a href="#Strains_Storage"><span class="tocnumber"></span> <span class="toctext">Strains Storage</span></a></li>
+
-
</ul>
+
-
</td></tr></tbody></table>
+
<br>
<br>
-
 
+
==== BL21DE3 ====
-
 
+
===== Genotype =====
-
 
+
*<i>E. coli B F- dcm ompT hsdS(rB- mB-) gal [malB+]K-12(λS) </i>
-
 
+
===== Genetical Characteristics =====
-
 
+
* Antibiotic resistance: none!
-
 
+
===== Reference =====
-
 
+
* <html><a href="http://www.merckmillipore.com/germany/life-science-research/bl21de3-glycerol-stock/EMD_BIO-69387/p_52yb.s1OQ1MAAAEj7Bt9.zLX">Merck Millipore</a></html>
-
 
+
-
 
+
-
                                <!-- Text body -->
+
-
<tbody>
+
-
<tr valign="top" align="left">
+
-
<td style="padding: 0pt 20px 0pt 0pt;" width="910px">
+
-
<font face="Verdana" size="-1">
+
-
 
+
-
                                <!-- Ab hier kannst du editieren -->
+
-
 
+
-
<h2><b><a name="Recipes"></a>Recipes</b></h2>
+
-
<p align="justify" style="line-height:1.6em">
+
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Media"></a>Media</b></h3><br>
+
-
Recipes for liquid and solid media for bacterial cultivation.
+
-
<ol>
+
-
<li><u>LB-Medium</u> </li>
+
-
<div style="text-indent:20px;">1% (w/v) bacto tryptone </div>
+
-
<div style="text-indent:20px;">0.5% (w/v) bacto yeast extract </div>
+
-
<div style="text-indent:20px;">1% (w/v) NaCl </div>
+
-
<div style="text-indent:20px;">if required: ad 1% (w/v) agar </div>
+
-
</div>
+
<br>
<br>
-
 
+
==== XL1 blue ====
-
<li><u>M9-Medium</u> </li>
+
===== Genotype =====
 +
*<i>endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44 F'[ ::Tn10 proAB+ lacIq Δ(lacZ)M15] hsdR17(rK- mK+) </i>
 +
===== Genetical Characteristics =====
 +
* Antibiotic resistance: none!
 +
===== Reference =====
 +
* <html><a href="http://www.merckmillipore.com/life-science-research/competent-cells-spotlight-main/c_CzSb.s1O1SAAAAEhSc8sgj.x?back=true&cid=bios-x-goog-1UKEN1277">Merck Millipore</a></html>
<br>
<br>
-
 
+
==== MG1655 ====
-
<li><u>0.3% Tryptone swimming agar</u> </li>
+
===== Genotype =====
-
<div style="text-indent:20px;">1% (w/v) bacto tryptone </div>
+
*<i>F-λ-, rph-1</i>
-
<div style="text-indent:20px;">0.5% (w/v) NaCl </div>
+
===== Genetical Characteristics =====
-
<div style="text-indent:20px;">0.3% (w/v) agar </div>
+
* Antibiotic resistance: none!
 +
===== Reference =====
 +
*<html><a href="http://cgsc.biology.yale.edu/Strain.php?ID=4837">Coli Genetic Stock Center</a></html>
<br>
<br>
-
 
+
==== RP437 ====
 +
===== Genotype =====
 +
*<i>F-, thr-1, araC14, leuB6(Am), fhuA31, lacY1, tsx-78, λ-, eda-50, hisG4(Oc), rfbC1?, rpsL136(strR), xylA5, mtl-1, metF159(Am), thiE1</i>
 +
===== Genetical Characteristics =====
 +
* Antibiotic resistance: none!
 +
===== Reference =====
 +
*<html><a href="http://cgsc.biology.yale.edu/Strain.php?ID=111960">Coli Genetic Stock Center</a></html>
<br>
<br>
-
</ol>
+
=== Mutated strains ===
 +
==== <i>∆tar</i> ====
 +
===== Genotype =====
 +
*<i>F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, Δtar-739::kan, rph-1, Δ(rhaD-rhaB)568, hsdR514</i>
 +
===== Genetical Characteristics =====
 +
* Antibiotic resistance: Kanamycin
 +
===== Reference =====
 +
*<html><a href="http://cgsc.biology.yale.edu/Strain.php?ID=107842">Coli Genetic Stock Center</a></html>
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Antibiotics"></a>Antibiotics</b></h3><br>
 
-
Stock solutions and concentrations of antibiotics.
 
-
<ol>
 
-
<li></li>
 
-
</ol>
 
-
<br>
 
<br>
<br>
 +
=== Storage ===
 +
Storage of <i>E. coli</i> strains - How to keep <i>E. coli</i> strains.
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Supplements"></a>Supplements</b></h3><br>
+
==== Glycerol Stock ====
-
Stock solutions and concentrations of supplements.
+
The glycerol stock is prepared as follow:
-
<ol>
+
* 900 µL over night culture in LB-liquid medium
-
<li></li>
+
* 900 µL 50 % sterile glycerol
-
</ol>
+
-
<br>
+
-
<br>
+
-
 
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Buffer_and_Solution"></a>Buffer and Solution</b></h3><br>
 
-
Recipes for stocks of buffers and solutions.
 
-
<ol>
 
-
<li> <b><u>5x M9 salts stock solution</u></b> </li>
 
-
Stir until dissolved; sterilize by autoclaving at 121°C. <br>
 
-
<table width="50%" border="1" bordercolor="#000000" cellpadding="0" cellspacing="2">
 
-
<tr>
 
-
  <th scope="col">Amount  </th>
 
-
  <th scope="col">Substance</th>
 
-
</tr>
 
-
<tr>
 
-
  <td>64 g</td>
 
-
  <td>Na<sub>2</sub>HPO<sub>4</sub> &times; 7 H<sub>2</sub>O</td>
 
-
</tr>
 
-
<tr>
 
-
  <td>15 g </td>
 
-
  <td>KH<sub>2</sub>PO<sub>4</sub> </td>
 
-
</tr>
 
-
<tr>
 
-
  <td>2.5 g</td>
 
-
  <td>NaCl</td>
 
-
</tr>
 
-
<tr>
 
-
  <td>5.0 g</td>
 
-
  <td>NH<sub>4</sub>Cl</td>
 
-
</tr>
 
-
<tr>
 
-
  <td>ad 1000 ml</td>
 
-
  <td>ddH<sub>2</sub>O </td>
 
-
</tr>
 
-
</table>
 
-
<br>
 
-
<br>
 
-
 
-
 
-
<li><b><u>CaCl<sub>2</sub> Buffer for competent cells </u></b></li>
 
-
Stir until dissolved; sterilize by sterile filtration. <br>
 
-
<table width="50%" border="1" bordercolor="#000000" cellpadding="0" cellspacing="2">
 
-
<tr>
 
-
  <th scope="col">Amount</th>
 
-
  <th scope="col">Substance</th>
 
-
  <th scope="col">Notes</th>
 
-
</tr>
 
-
<tr>
 
-
  <td>7.351 g </td>
 
-
  <td>CaCl<sub>2</sub> &times; 7 H<sub>2</sub>O</td>
 
-
  <td>Alternative 0.1 M CaCl<sub>2</sub>.</td>
 
-
</tr>
 
-
<tr>
 
-
  <td>15%        </td>
 
-
  <td> Glycerol</td>
 
-
  <td></td>
 
-
</tr>
 
-
  <tr>
 
-
  <td>ad 500 ml</td>
 
-
  <td>ddH<sub>2</sub>O</td>
 
-
  <td></td>
 
-
</tr>
 
-
</table>
 
-
<br>
 
-
<br>
 
-
 
-
<li><b><u> 50x TAE for agarose gels</u></b></li>
 
-
Stir until dissolved. Working concentration is 1x (40 ml of 50x stock ad 2 l ddH<sub>2</sub>O).<br>
 
-
<table width="50%" border="1" bordercolor="#000000" cellpadding="0" cellspacing="2">
 
-
<tr>
 
-
  <th scope="col">Amount</th>
 
-
  <th scope="col">Substance</th>
 
-
</tr>
 
-
<tr>
 
-
  <td>242 g</td>
 
-
  <td>Tris base</td>
 
-
</tr>
 
-
<tr>
 
-
  <td>57.1 ml        </td>
 
-
  <td>Glacial acetic acid  </td>
 
-
</tr>
 
-
  <tr>
 
-
  <td>18.6 g</td>
 
-
  <td>EDTA </td>
 
-
</tr>
 
-
  <tr>
 
-
  <td>ad 1000 ml  </td>
 
-
  <td>ddH<sub>2</sub>O </td>
 
-
</tr>
 
-
</table>
 
-
</ol>
 
-
<br>
 
-
<br>
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
<h2><b><a name="E._coli"></a><i>E. coli</i></b></h2>
 
-
<p align="justify" style="line-height:1.6em">
 
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Strains"></a>Strains</b></h3><br>
 
-
<i>E. coli</i> strain list - List of the working <i>E. coli</i> strains including genotypes, genetic markers, and alleles.
 
-
<ol>
 
-
<li></li>
 
-
</ol>
 
-
<br>
 
-
<br>
 
-
 
-
 
-
<h3 style="margin:3px; background:#cedff2; font-size:120%; font-weight:bold; border:1px solid #a3b0bf; text-align:left; color:#000; padding:0.2em 0.4em;"<b><a name="Strains_Storage"></a>Storage</b></h3><br>
 
-
Storage of <i>E. coli</i> strains - How to keep <i>E. coli</i> strains. </li>
 
-
<ol>
 
-
<li></h3><b><u>Glycerol Stock</u></b></li>
 
-
<ul>
 
-
<li>900 µl over night culture in LB-liquid medium </li>
 
-
<li>900 µl 50% sterile glycerol</li>
 
-
</ul>
 
Glycerol is extremely viscous. Hence, use cut off tips to pipette. Glycerol stocks were vortexed to ensure uniformly mixture and shock
Glycerol is extremely viscous. Hence, use cut off tips to pipette. Glycerol stocks were vortexed to ensure uniformly mixture and shock
-
frozen in N<sub>2</sub> or dry ice. Then, cultures were stored at -80°C. Do NOT forget to fill in the according GMO S1 construction sheet!
+
frozen in N<sub>2</sub> or dry ice. Then, cultures were stored at -80 °C. Do NOT forget to fill in the according GMO S1 construction sheet!
-
</ol>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<br>
+
-
<br>
+
-
<body id="top">
+
-
<a href="#top">&uarr; Return to top</a>
+
-
<br>
+
-
<br>
+
-
 
+
-
 
+
-
 
+
-
<!-- sponsorbar -->
+
-
<br><br>
+
-
<table width=900>
+
-
<tr><td>
+
-
 
+
-
<hr>
+
-
<h2><span><b>Team G&ouml;ttingen Sponsors and Supporter</b></span></h2>
+
-
</td></tr>
+
-
</table>
+
-
 
+
-
<table width=900>
+
-
    <tr>
+
-
          <td><center><u>Head Sponsor:</u></center></td><td width=1></td>
+
-
          <td><center><u>Premium Sponsor:</u></center></td><td width=1></td>
+
-
          <td><center><u>Supporter:</u></center></td>
+
-
    </tr>
+
-
 
+
-
    <tr>
+
-
          <td><a href="http://www.kws.com"><img width="150 px" src="http://www.patrickreinke.de/igem/kws.png"></a></td><td width=1; bgcolor=grey></td>
+
-
          <td><a href="http://www.sartorius.com"><img width="250 px" src="http://www.patrickreinke.de/igem/sartorius.png"></a></td><td width=1; bgcolor=grey></td>
+
-
          <td>
+
-
              <img width="150 px" src="http://www.patrickreinke.de/igem/mpi.jpg">
+
-
              <img width="150 px" src="http://www.patrickreinke.de/igem/gzmb.jpg">
+
-
              <img width="150 px" src="http://www.patrickreinke.de/igem/eppendorf.jpg">
+
-
          </td>
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
<!-- sponsorbar -->
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
 +
==== LB-agar ====
 +
LB-agar plates were prepared as follow:
 +
* 1% agar (w/v) was added to 1 L LB-broth
 +
* LB-agar was autoclaved
 +
* LB-agar was poured in plates
-
                                <!-- Text body zu Ende -->
+
The <i>E. coli</i> strains were plated out and stored at 4°C. The plates were renewed every two weeks. 
-
</font></td></tr></tbody></table></div>
+
{{GoettingenFooter}}
-
</td></tr>
+
sub>2
-
</table>
+
-
</body>
+
-
</html>
+

Latest revision as of 16:15, 26 September 2012

Deutsch  / English 

Contents

Recipes

Media

Recipes for liquid and solid media for bacterial cultivation. Stir until dissolved; sterilize by autoclaving 121 °C. Do not forget a magnetic stirrer.

LB-broth

  • 1 % (w/v) bacto tryptone
  • 0.5 % (w/v) bacto yeast extract
  • 1 % (w/v) NaCl
  • if required: ad 1 % (w/v) agar

M9 Swimming Agar

  • 1.25 % (v/v) glycerol
  • 20 % (v/v) 5x M9 salt stock solution
  • 0.1 % (v/v) of CaCl2 x 2 H2O stock solution (20 mg/mL)
  • 0.1 % (v/v) MgSO4 stock solution (0.12 g/mL)
  • 0.3 % (w/v) of agar

Tryptone swimming agar

  • 1 % (w/v) bacto tryptone
  • 0.5 % (w/v) NaCl
  • 0.3 % (w/v) agar

Antibiotics

  • Stock solutions and concentrations of antibiotics.
Reagent Abbreviation Stock Conc. [mg/mL] Working Conc. [µg/mL] Dilution Solvent Notes
Ampicillin Amp 100 100 1,000x ddH2O Culture plates with Amp can be stored at 4 °C for about 2 weeks. Stock solutions can be stored at 4 °C for 2 weeks but can last as long as 4-6 months when stored at -20 °C.
Chloramphenicol Cm 20 20 1,000x EtOH
Kanamycin Kan 50 50 1,000x ddH2O Plates may only last a few weeks at 4 °C.


Supplements

  • Aminoacid mix (containing all aminoacids except tryptophane): stocksolution 0.2 % (w/v), varying amounts used in swimming assays, view methods and notebook of group 1
  • Caffeine: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • D-Aspartate: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • 2-Ethyl-1-Hexanole: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • Geraniol: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • L-Aspartate-4-Benzylester: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • L-aspartate: stocksolution 10 mM, varying amounts used in swimming assays, view methods and notebook of group 1
  • Leucin: stocksolution 30.49 mM, add to M9-agar: 4 mL (final concentration 0.305 µM)
  • Methionin: stocksolution 26.8 mM, add to M9-agar: 4 mL (final concentration 0.268 µM)
  • Sodium Cyclamate: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
  • Tryptone: stocksolution 0.5 % (w/v), varying amounts used in swimming assays, view methods and notebook of group 1
  • Vanillin: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods

Buffer and Solution

Recipes for stocks of buffers and solutions.

5x M9 salts stock solution

Amount Substance
64 g Na2HPO4 × 7 H2O
15 g KH2PO4
2.5 g NaCl
5.0 g NH4Cl
ad 1000 ml ddH2O
  • Stir until dissolved

CaCl2 Buffer for competent cells

Amount Substance Notes
7.351 g CaCl2 × 7 H2O Alternative 0.1 M CaCl2
15% Glycerol
ad 500 ml ddH2O
  • Stir until dissolved; sterilize by sterile filtration.

50x TAE for agarose gels

Amount Substance
242 g Tris base
57.1 mL Glacial acetic acid
18.6 g EDTA
ad 1000 mL ddH2O
  • Stir until dissolved. Working concentration is 1x (40 mL of 50x stock ad 2 L ddH2O).

E. coli

  • E. coli strain list - List of the working E. coli strains including genotypes, genetic markers, and alleles.

Non-mutated strains

These strains may not contain any plasmids or resistances, because they should be used for transformation or other experiments.

DH10B

Genotype
  • F- endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74 Φ80lacZΔM15 araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) λ-
Genetical Characteristics
  • Antibiotic resistance: none
Reference


DH5α

Genotype
  • F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ–
Genetical Characteristics
  • Antibiotic resistance: none
Reference


BL21DE3

Genotype
  • E. coli B F- dcm ompT hsdS(rB- mB-) gal [malB+]K-12(λS)
Genetical Characteristics
  • Antibiotic resistance: none!
Reference


XL1 blue

Genotype
  • endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44 F'[ ::Tn10 proAB+ lacIq Δ(lacZ)M15] hsdR17(rK- mK+)
Genetical Characteristics
  • Antibiotic resistance: none!
Reference


MG1655

Genotype
  • F-λ-, rph-1
Genetical Characteristics
  • Antibiotic resistance: none!
Reference


RP437

Genotype
  • F-, thr-1, araC14, leuB6(Am), fhuA31, lacY1, tsx-78, λ-, eda-50, hisG4(Oc), rfbC1?, rpsL136(strR), xylA5, mtl-1, metF159(Am), thiE1
Genetical Characteristics
  • Antibiotic resistance: none!
Reference


Mutated strains

∆tar

Genotype
  • F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, Δtar-739::kan, rph-1, Δ(rhaD-rhaB)568, hsdR514
Genetical Characteristics
  • Antibiotic resistance: Kanamycin
Reference


Storage

Storage of E. coli strains - How to keep E. coli strains.

Glycerol Stock

The glycerol stock is prepared as follow:

  • 900 µL over night culture in LB-liquid medium
  • 900 µL 50 % sterile glycerol

Glycerol is extremely viscous. Hence, use cut off tips to pipette. Glycerol stocks were vortexed to ensure uniformly mixture and shock frozen in N2 or dry ice. Then, cultures were stored at -80 °C. Do NOT forget to fill in the according GMO S1 construction sheet!

LB-agar

LB-agar plates were prepared as follow:

  • 1% agar (w/v) was added to 1 L LB-broth
  • LB-agar was autoclaved
  • LB-agar was poured in plates

The E. coli strains were plated out and stored at 4°C. The plates were renewed every two weeks.

↑ Back to top!
sub>2

Retrieved from "http://2012.igem.org/Team:Goettingen/Project/Materials"