Team:Heidelberg/Notebook/Sensing Group/Notebook/2ndweek
From 2008.igem.org
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Chenchenzhu (Talk | contribs) (→Wednesday, 08/13/2008) |
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<html> | <html> | ||
- | <link | + | |
- | + | ||
- | + | <style> | |
+ | h1.firstHeading { display: none; } | ||
+ | |||
+ | p {text-align: justify;} | ||
+ | |||
+ | a:link { color: #00b0e6; text-decoration: none} | ||
+ | a:visited { color:#00b0e6; text-decoration: none} | ||
+ | a:hover { color:#f29400; text-decoration: none} | ||
+ | a:active { color:#f29400; text-decoration: none} | ||
+ | |||
+ | #bodyContent { padding: 10px auto; width: 910px; margin: auto; clear: none; } | ||
+ | |||
+ | table#team_members { text-align: justify; border: 0; } | ||
+ | table#team_members h2, table#team_members h3 { clear: both; } | ||
+ | |||
+ | |||
+ | /*-----------------------------------------------------------------------------------------------*/ | ||
+ | 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: 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 */ | ||
+ | * 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: #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; | ||
+ | } | ||
+ | |||
+ | |||
+ | 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: #00b0e6; /* 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. */ | ||
+ | } | ||
+ | /*...............................................................................................*/ | ||
+ | |||
+ | </style> | ||
+ | |||
<body> | <body> | ||
Line 52: | Line 393: | ||
</li> | </li> | ||
<li> | <li> | ||
- | <a href="https://2008.igem.org/Team:Heidelberg/ | + | <a href="https://2008.igem.org/Team:Heidelberg/Modeling" style="color: white">Modeling<!--[if gt IE 6]><!--></a><!--<![endif]--> |
<!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]--> | <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]--> | ||
<ul class="DropDownMenu" id="MB1-DDM3"> | <ul class="DropDownMenu" id="MB1-DDM3"> | ||
Line 62: | Line 403: | ||
</li> | </li> | ||
<li> | <li> | ||
- | <a href="https://2008.igem.org/Team:Heidelberg/Notebook/ | + | <a href="https://2008.igem.org/Team:Heidelberg/Notebook/Overview" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]--> |
<!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]--> | <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]--> | ||
<ul class="DropDownMenu" id="MB1-DDM5"> | <ul class="DropDownMenu" id="MB1-DDM5"> | ||
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</li> | </li> | ||
<li style="width: 160px"> | <li style="width: 160px"> | ||
- | <a href="https://2008.igem.org/Team:Heidelberg/ | + | <a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Project_Overview" 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]--> | <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]--> | ||
<ul class="DropDownMenu" id="MB1-DDM4"> | <ul class="DropDownMenu" id="MB1-DDM4"> | ||
Line 106: | Line 447: | ||
<li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Surveys"><span><span>Surveys</span></span></a></li> | <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Surveys"><span><span>Surveys</span></span></a></li> | ||
<li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Open_Day"><span><span>Open Day</span></span></a></li> | <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Open_Day"><span><span>Open Day</span></span></a></li> | ||
- | <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Nobel_Prize"><span><span>Nobel Prize</span></span></a></li> | + | <li><a href="https://2008.igem.org/Team:Heidelberg/Human_Practice/Nobel_Prize"><span><span>Nobel Prize</span></span></a></li> |
+ | |||
</ul> | </ul> | ||
<!--[if lte IE 6]></td></tr></table></a><![endif]--> | <!--[if lte IE 6]></td></tr></table></a><![endif]--> | ||
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== Tuesday, 08/12/2008 == | == Tuesday, 08/12/2008 == | ||
* Miniprep of correctly sequenced LuxS and Glycerol-stock | * Miniprep of correctly sequenced LuxS and Glycerol-stock | ||
- | * LuxQ no. 3 checked via digestion with NcoI/BamHI --> negative result | + | * LuxQ no. 3 checked via digestion with NcoI/BamHI (NEBuffer 3 + BSA) --> negative result |
* PCR for LuxQ, LuxQ-1, LuxQ-2, Tar-1, Tar-2 with Taq Mastermix | * PCR for LuxQ, LuxQ-1, LuxQ-2, Tar-1, Tar-2 with Taq Mastermix | ||
** 5min @ 95°C || 30s @ 95°C | 30s @ 58 °C | 2min @ 72°C || 10min @ 72°C | 4°C hold (30 cycles) | ** 5min @ 95°C || 30s @ 95°C | 30s @ 58 °C | 2min @ 72°C || 10min @ 72°C | 4°C hold (30 cycles) | ||
Line 135: | Line 477: | ||
== Wednesday, 08/13/2008 == | == Wednesday, 08/13/2008 == | ||
* PCR-purification of the all PCR-products eluted in 50 µl H<sub>2</sub>O | * PCR-purification of the all PCR-products eluted in 50 µl H<sub>2</sub>O | ||
- | * Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI | + | * Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI (NEBuffer 2 + BSA) |
* Fusion-PCR for Fusion-1 and repetition of PCR for LuxQ-2 and LuxQ | * Fusion-PCR for Fusion-1 and repetition of PCR for LuxQ-2 and LuxQ | ||
* digestion of pDK48 plasmid and Fusion-1 frament with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4 | * digestion of pDK48 plasmid and Fusion-1 frament with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4 | ||
- | + | [[Image:HD 080813-LuxQ PCR-pDK48 mini LuxQ3 xbaI.png|left|thumb|500px|Fusion-PCR for Fusion-1, LuxQ PCR, LuxQ-2 PCR, pDK48 miniprep and xbaI digestion of LuxQ no. 3]] [[Image:HD 080813-F1 pDK48 Digestion.png|left|thumb|200px|Digestion of Fusion-1 and pDK48 with NcoI/NdeI/AgeI]] | |
- | + | ||
<br style="clear: both;"/> | <br style="clear: both;"/> | ||
Line 145: | Line 486: | ||
== Thursday, 08/14/2008 == | == Thursday, 08/14/2008 == | ||
[[Image:HD 080814-LuxQ PCR.png|thumb|right|PCR for LuxQ and LuxQ-2]] | [[Image:HD 080814-LuxQ PCR.png|thumb|right|PCR for LuxQ and LuxQ-2]] | ||
- | * Gel | + | * Gel for LuxQ, LuxQ2 |
* sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3) | * sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3) | ||
* Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4) | * Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4) | ||
Line 153: | Line 494: | ||
== Friday, 08/15/2008 == | == Friday, 08/15/2008 == | ||
* digestion of Fusion-1 with AgeI in NEBuffer 1 | * digestion of Fusion-1 with AgeI in NEBuffer 1 | ||
- | * | + | * gel extraction of sequentially digested Fusion-1 and pDK48 products |
- | * new digestion of Fusion-1 with NcoI/NdeI and | + | * new digestion of Fusion-1 with NcoI/NdeI (NEBuffer 4) and gel extraction, because previous digestion did not yield expected bands (there should be two small bands at 516bp and 392bp) |
[[Image:HD 080815-F1 digestion pDK48.png|left|thumb|250px|Gelextraction of digested Fusion-1 and purified PCR product as well as pDK48 digested. Expected bands at 516bp and 392bp are not visible.]] | [[Image:HD 080815-F1 digestion pDK48.png|left|thumb|250px|Gelextraction of digested Fusion-1 and purified PCR product as well as pDK48 digested. Expected bands at 516bp and 392bp are not visible.]] | ||
[[Image:HD 080815-F1 digestion NcoI NdeI.png|left|thumb|200px|Fusion-1 digested with NcoI/NdeI. Expected bands of 1025bp and 908bp had to be cut out together. Wanted band is the longer one.]] | [[Image:HD 080815-F1 digestion NcoI NdeI.png|left|thumb|200px|Fusion-1 digested with NcoI/NdeI. Expected bands of 1025bp and 908bp had to be cut out together. Wanted band is the longer one.]] | ||
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<br style="clear: both;"/> | <br style="clear: both;"/> | ||
- | * Ligation of 5 µl Fusion-1 | + | * Ligation of 5 µl Fusion-1 with 5 µl pDK48 (both digested with NcoI/NdeI ) |
- | * Transformation into | + | * Transformation into DH5α (5 µl Ligation products) |
[[Team:Heidelberg/Notebook/Sensing_Group/Notebook/3rdweek | Go to 3rd week]] | [[Team:Heidelberg/Notebook/Sensing_Group/Notebook/3rdweek | Go to 3rd week]] |
Latest revision as of 14:02, 29 October 2008
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Contents |
Monday, 08/11/2008
- preparation of LuxS harboring cells O/N culture
Tuesday, 08/12/2008
- Miniprep of correctly sequenced LuxS and Glycerol-stock
- LuxQ no. 3 checked via digestion with NcoI/BamHI (NEBuffer 3 + BSA) --> negative result
- PCR for LuxQ, LuxQ-1, LuxQ-2, Tar-1, Tar-2 with Taq Mastermix
- 5min @ 95°C || 30s @ 95°C | 30s @ 58 °C | 2min @ 72°C || 10min @ 72°C | 4°C hold (30 cycles)
- preparation of O/N culture for LuxQ no. 3
Wednesday, 08/13/2008
- PCR-purification of the all PCR-products eluted in 50 µl H2O
- Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI (NEBuffer 2 + BSA)
- Fusion-PCR for Fusion-1 and repetition of PCR for LuxQ-2 and LuxQ
- digestion of pDK48 plasmid and Fusion-1 frament with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4
Thursday, 08/14/2008
- Gel for LuxQ, LuxQ2
- sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3)
- Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
Friday, 08/15/2008
- digestion of Fusion-1 with AgeI in NEBuffer 1
- gel extraction of sequentially digested Fusion-1 and pDK48 products
- new digestion of Fusion-1 with NcoI/NdeI (NEBuffer 4) and gel extraction, because previous digestion did not yield expected bands (there should be two small bands at 516bp and 392bp)
- Ligation of 5 µl Fusion-1 with 5 µl pDK48 (both digested with NcoI/NdeI )
- Transformation into DH5α (5 µl Ligation products)