Team:UQ-Australia/Notebook/Transformation procedure2

__NOTOC__
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li:hover works in IE7 and FF2.
a:hover works in IE6 and FF2.
a:hover breaks li:hover in FF2.
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div.MenuBar ul li a:hover ul.DropDownMenu li a ul.SideMenu {
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}

/*------------------------------------------------------------------------------ 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: 23px; /* 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: black; /* Selected item. */
  color: #FFFFFF;

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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: black; /* 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. */
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  height: 20px;
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}
/*----------------------------------------------------------------------------------- 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 arial, helvetica, sans-serif; /* 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 arial, helvetica, sans-serif; /* 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. */
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* html ul.DropDownMenu li a:hover {
  cursor: hand; /* Required for IE55.
                   Hidden (* html) from non-IE browsers. */
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ul.DropDownMenu li a:hover {
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                      Hidding it (* html) from non-IE browsers breaks IE7! 
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* html div.MenuBar a:hover {
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                            Hidden (* html) from non-IE browsers. */
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* html div.MenuBar ul li table,
* html div.MenuBar ul li table td {
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                Hidden (* html) from non-IE browsers. */
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div.MenuBar a {
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  color: MenuText;
}
div.MenuBar ul li:hover a,
div.MenuBar ul li a:hover {
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  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; 
}
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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: black; /* 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: black; /* 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: #603371; /* 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 {
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}
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: 12em; /* 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: 10em; /* 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: 10em; /* 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: 10em; /* Side menu width. */
}
/*...............................................................................................*/

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==Transformation Procedure 2==

1. Thaw competent cells on ice. Place number of required polypropylene tubes on ice. 

2. Gently mix cells, then aliquot 100ul competent cells into chilled polypropylene tubes. Add 1.7 ul of B-mercaptoethanol. Gently stir every 2 minutes for 10 minutes.

3. Add 1-10ng of plasmid DNA, moving the pipette through cells while dispersing. Gently tap tubes to mix. 

4. Incubate cells on ice for 30 minutes

5. Heat-shock cells for 30 seconds in 42'C waterbath. DO NOT SHAKE

6. Place on ice for 2 minutes

7. Add 900ul of room temperature SOC medium + glucose

8. Shake at 225rpm (37'C) for 1 hour

9. During the 1 hour incubation take agar plates out of fridge and incubate in 37'C incubator for 30 minutes

10. Spread 3 agar plates: 50ul, 100ul, 500ul

11. Incubate at 37'C overnight