Team:UQ-Australia/Notebook/Transformation procedure2



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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. 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: arial, helvetica, sans-serif; height: 30px; width: 910px; /*width: 100%*/ margin: 0; border-top: 0; border-right: 0; border-left: 0; padding: 0; background: black; } div.MenuBar ul { font: arial, helvetica, sans-serif; text-align: center; list-style-type: none; margin: 0.5em auto; border: 0; padding: 0; background: black; } div.MenuBar ul li { font: arial, helvetica, sans-serif; display: block; padding: 0; margin: 0; font-size: 1.3em; float: left; background: black; text-align: center; width: 107px; position: relative; /* Sets the positioning context for each drop-down menu. */ }

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; }
 * html div.MenuBar ul li a {
 * html ul.DropDownMenu li a:hover {
 * 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 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