Team:Washington/OVERNIGHT CULTURES
From 2013.igem.org
(Created page with "=="""== <html> <body> <h1>Preparation of Overnight Cultures</h1> <p>Use TB for making glycerol stocks and TB for minipreps.</p> <ol> <li>Obtain a 14 mL Sterile Polypropylen...") |
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+ | |||
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+ | position: absolute; | ||
+ | top: 40px; | ||
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+ | z-index: 1; | ||
+ | background-color: #444; | ||
+ | display: none; | ||
+ | -moz-box-shadow: none; | ||
+ | -webkit-box-shadow: none; | ||
+ | box-shadow: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu:after { | ||
+ | content: ''; | ||
+ | position: absolute; | ||
+ | left: 25px; | ||
+ | top: -8px; | ||
+ | border-left: 8px solid transparent; | ||
+ | border-right: 8px solid transparent; | ||
+ | border-bottom: 8px solid #444; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul { | ||
+ | position: static; | ||
+ | visibility: visible; | ||
+ | opacity: 1; | ||
+ | margin: 0; | ||
+ | background: none; | ||
+ | -moz-box-shadow: none; | ||
+ | -webkit-box-shadow: none; | ||
+ | box-shadow: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul ul { | ||
+ | margin: 0 0 0 20px !important; | ||
+ | -moz-box-shadow: none; | ||
+ | -webkit-box-shadow: none; | ||
+ | box-shadow: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu li { | ||
+ | position: static; | ||
+ | display: block; | ||
+ | float: none; | ||
+ | border: 0; | ||
+ | margin: 5px; | ||
+ | -moz-box-shadow: none; | ||
+ | -webkit-box-shadow: none; | ||
+ | box-shadow: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul li{ | ||
+ | margin-left: 20px; | ||
+ | -moz-box-shadow: none; | ||
+ | -webkit-box-shadow: none; | ||
+ | box-shadow: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu a{ | ||
+ | display: block; | ||
+ | float: none; | ||
+ | padding: 0; | ||
+ | color: #999; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu a:hover{ | ||
+ | color: #fafafa; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul a{ | ||
+ | padding: 0; | ||
+ | width: auto; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul a:hover{ | ||
+ | background: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu ul li:first-child a:after, | ||
+ | #igemuwmenu ul ul li:first-child a:after { | ||
+ | border: 0; | ||
+ | } | ||
+ | |||
+ | } | ||
+ | |||
+ | @media screen and (min-width: 600px) { | ||
+ | #igemuwmenu { | ||
+ | display: block !important; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | /* iPad */ | ||
+ | .no-transition { | ||
+ | -webkit-transition: none; | ||
+ | -moz-transition: none; | ||
+ | -ms-transition: none; | ||
+ | -o-transition: none; | ||
+ | transition: none; | ||
+ | opacity: 1; | ||
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+ | display: none; | ||
+ | } | ||
+ | |||
+ | #igemuwmenu li:hover > .no-transition { | ||
+ | display: block; | ||
+ | } | ||
+ | |||
+ | </style> | ||
+ | |||
+ | <a href="https://2013.igem.org/Team:Washington"><img class="thumbnail" src='https://static.igem.org/mediawiki/2013/1/1b/Team_Washington2013_banner_final.jpg' width= "965"/></A> | ||
+ | |||
+ | <a href="http://www.washington.edu/"><img src='https://static.igem.org/mediawiki/2012/2/25/UW_W-Logo_smallRGB.jpg' align="left" width= "57" style="padding-top: 12px; padding-left: 5px;"/></a> | ||
+ | |||
+ | <a href="https://2012.igem.org/Main_Page"><img src='https://static.igem.org/mediawiki/igem.org/c/c8/IGEM_desat.png' align="right" width= "70"/></a> | ||
+ | |||
+ | <style type="text/css"> | ||
+ | #content {z-index:4;} | ||
+ | .expand {background: url('https://static.igem.org/mediawiki/2008/e/ef/Icon-expand.png') no-repeat 95% 50%;} | ||
+ | .collapse {background: url('https://static.igem.org/mediawiki/2008/c/cd/Icon-collapse.png') no-repeat 95% 50%;} | ||
+ | .docked {background: #99ccff url("https://static.igem.org/mediawiki/2008/6/62/Ddnavundock.png") no-repeat 50% 50%;} | ||
+ | .undocked {background: #99ccff url("https://static.igem.org/mediawiki/2008/e/e4/Ddnavdock.png") no-repeat 50% 50%;} | ||
+ | </style> | ||
+ | |||
+ | <nav id="igemuwmenu-wrap"> | ||
+ | |||
+ | <ul id="igemuwmenu"> | ||
+ | |||
+ | <li><a href="https://2013.igem.org/Team:Washington">Home</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington">UW 2013</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Washington">UW 2012</a></li> | ||
+ | <li><a href="https://2011.igem.org/Team:Washington">UW 2011</a></li> | ||
+ | <li><a href="https://2010.igem.org/Team:Washington">UW 2010</a></li> | ||
+ | <li><a href="https://2009.igem.org/Team:Washington">UW 2009</a></li> | ||
+ | <li><a href="https://2008.igem.org/Team:University_of_Washington">UW 2008</a></li> | ||
+ | <li><a href="https://2013.igem.org/Main_Page">iGEM Homepage</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | <!-- | ||
+ | <li> | ||
+ | <a </a> main menu on plastic degradation | ||
+ | <ul> | ||
+ | |||
+ | for the drop down of plastic degradation | ||
+ | format <li><a href="https://2012.igem.org/Team:Washington/Plastics#Background">Background</a></li> | ||
+ | |||
+ | |||
+ | </ul> | ||
+ | </li> | ||
+ | --> | ||
+ | |||
+ | |||
+ | <li> | ||
+ | <a href="https://2013.igem.org/Team:Washington/LightSensing">Light Sensing Experiments</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Background">Background</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Cloning">Cloning</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Systems">Our Systems</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Methods">Methods</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Results">Results</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Sources">Scientific Sources</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Future">Future Plans</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/LightSensing#Parts">Parts Submitted</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | |||
+ | <li> | ||
+ | <a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE">e.colightTune</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Background">Background</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Usage">App</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Benefits">Methods</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Results">Results Summary</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | <li> | ||
+ | <a href="https://2013.igem.org/Team:Washington/Outreach">Outreach</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/Outreach#Activities">Activities</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/Outreach#Pictures">Pictures</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | <li> | ||
+ | <a href="https://2013.igem.org/Team:Washington/Protocols">Protocols</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/Protocols#General_Protocols">General</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/Protocols#Cloning_Protocols">Cloning</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/Protocols#Assay">Light assays</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | <li> | ||
+ | <a href="https://2013.igem.org/Team:Washington/MoreLinks">Links</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/MoreLinks#Safety">Safety Form</a></li> | ||
+ | <li><a href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2012&group=Washington">Parts Registry</a></li> | ||
+ | <li><a href="https://2013.igem.org/Team:Washington/MoreLinks#Documentation">Parts Documentation</a></li> | ||
+ | |||
+ | </ul> | ||
+ | </li> | ||
+ | |||
+ | <li style="border-right:0; box-shadow:0px 0 0;"> | ||
+ | <a href="https://2013.igem.org/Team:Washington/Funding">Funding</a> | ||
+ | </li> | ||
+ | |||
+ | </li> | ||
+ | |||
+ | </ul> | ||
+ | </nav> | ||
+ | |||
+ | <script type="text/javascript" src="http://code.jquery.com/jquery-latest.min.js"></script> | ||
+ | <script type="text/javascript"> | ||
+ | $(function() { | ||
+ | if ($.browser.msie && $.browser.version.substr(0,1)<7) | ||
+ | { | ||
+ | $('li').has('ul').mouseover(function(){ | ||
+ | $(this).children('ul').css('visibility','visible'); | ||
+ | }).mouseout(function(){ | ||
+ | $(this).children('ul').css('visibility','hidden'); | ||
+ | }) | ||
+ | } | ||
+ | |||
+ | /* Mobile */ | ||
+ | $('#igemuwmenu-wrap').prepend('<div id="igemuwmenu-trigger">igemuwmenu</div>'); | ||
+ | $("#igemuwmenu-trigger").on("click", function(){ | ||
+ | $("#menu").slideToggle(); | ||
+ | }); | ||
+ | |||
+ | // iPad | ||
+ | var isiPad = navigator.userAgent.match(/iPad/i) != null; | ||
+ | if (isiPad) $('#igemuwmenu ul').addClass('no-transition'); | ||
+ | }); | ||
+ | // selection | ||
+ | |||
+ | </script> | ||
+ | |||
+ | <script type="text/javascript"> | ||
+ | $("#content").find("a[href='"+location.href+"']").each(function(){ | ||
+ | $(this).find("a").addClass("selected") | ||
+ | }) | ||
+ | </script> | ||
+ | |||
+ | <script type="text/javascript"> | ||
+ | |||
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+ | |||
+ | |||
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<p>1. Label a 15 mL culture tube with strain/gene, antibiotic, and date.<br /> | <p>1. Label a 15 mL culture tube with strain/gene, antibiotic, and date.<br /> | ||
2. Add the following to tube as appropriate:</p> | 2. Add the following to tube as appropriate:</p> | ||
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Latest revision as of 07:32, 7 September 2013
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Preparation of Overnight Cultures
Use TB for making glycerol stocks and TB for minipreps.
- Obtain a 14 mL Sterile Polypropylene Round-Bottom Tube.
- Add ~ 3-5 mLs of an appropriate broth (either LB or TB)* see tips
- Add an appropriate antibiotic if necessary and set this tube safely aside
- Amp (1:1000)
- Chlor (1:1000)
- Kan (1:1000)
- Incubate overnight cultures @ 37oC, overnight in the iGEM labeled shaker
General Purpose Short Protocol:
Set-up Time: 5 min
Total Time: 12 - 24 hours (18 hours for best plasmid production)
1. Label a 15 mL culture tube with strain/gene, antibiotic, and date.
2. Add the following to tube as appropriate:
Reagent |
Amount |
Where to Find it |
Broth (LB or TB) |
3-5 mL (as needed) |
Room temp or fridge stocks |
Antibiotic(s) |
1 uL/mL - Amp/Carb |
-20 C Freezer or Refrigerator |
Cells |
1 colony |
Refrigerator |
3. Gently mix, and place in 37C shaker-incubator overnight, with lid attached loosely for good aeration.
*Broth:
The type of broth used varies by the application. If we are growing up cells to miniprep, we typically use TB to get a higher OD of cells, though LB works well for any other applications. Broth that is pre-mixed with antibiotics should be stored in the refrigerator for no longer than 2 months.
2013 UW iGEM is using LB from capsules which has gelatin that will clog purification columns. The TB should be made from powder.