Team:UQ-Australia/Notebook/Transformation procedure



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div.MenuBar ul li a { font: arial, helvetica, sans-serif; display: block; background: black; height: 22px; /* Keep height + padding-top + padding-bottom sync with the menu bar height. */ color: #ffffff; 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: 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;

} 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. */ 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 arial, helvetica, sans-serif; /* 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 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 */ float: left; /* Required for IE55 and IE6. Breaks O9. Hidden (* html) from non-IE browsers. */ } 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! } text-decoration: none; /* Required for IE55 and IE6. Hidden (* html) from non-IE browsers. */ } 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: #ea7f16; 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; } /*---*/ /*Script-Free 3-Level Menu 1.2 Tailor www.CesarDaniel.info /*-- General */ body { background: white; color: black; margin: 0; border: 0; padding: 0; }
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 * html div.MenuBar a:hover {
 * html div.MenuBar ul li table,
 * html div.MenuBar ul li table td {

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 { width: 11em; /* 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: 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
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.

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

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