Difference between revisions of "Team:UMaryland/protocols"
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background-size: 1000% ; | background-size: 1000% ; | ||
background-repeat: no-repeat; | background-repeat: no-repeat; | ||
+ | } | ||
+ | #sidemenu { | ||
+ | position:fixed; | ||
+ | float:left; | ||
+ | width:200px; | ||
+ | top:200px; | ||
+ | z-index:2; | ||
} | } | ||
− | # | + | #sidemenu, |
− | + | #sidemenu ul, | |
− | + | #sidemenu ul li, | |
− | + | #sidemenu ul li a { | |
− | + | margin: 0; | |
− | + | padding: 0; | |
− | + | border: 0; | |
− | + | list-style: none; | |
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+ | display: block; | ||
+ | -webkit-box-sizing: border-box; | ||
+ | -moz-box-sizing: border-box; | ||
+ | box-sizing: border-box; | ||
} | } | ||
− | # | + | |
− | + | #sidemenu ul ul { | |
− | + | display: none; | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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} | } | ||
− | + | .align-right { | |
− | + | float: right; | |
− | + | ||
− | + | ||
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} | } | ||
− | # | + | #sidemenu > ul > li > a { |
− | + | padding: 15px 20px; | |
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+ | cursor: pointer; | ||
+ | z-index: 2; | ||
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+ | box-shadow: inset 0 1px 0 rgba(255, 255, 255, 0.15); | ||
+ | } | ||
+ | |||
+ | #sidemenu > ul > li > a:hover, | ||
+ | #sidemenu > ul > li.active > a, | ||
+ | #sidemenu > ul > li.open > a { | ||
+ | color: #000000; | ||
+ | background-image: -webkit-linear-gradient(top, #C9C9C9 0%, #FFFFFF 100%); | ||
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+ | #sidemenu > ul > li.open > a { | ||
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+ | border-bottom: 1px solid #1682ba; | ||
+ | } | ||
+ | |||
+ | |||
+ | #sidemenu > ul > li > a:hover > span::after, | ||
+ | #sidemenu > ul > li.active > a > span::after, | ||
+ | #sidemenu > ul > li.open > a > span::after { | ||
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+ | box-shadow: inset 0 1px 0 rgba(255, 255, 255, 0.1); | ||
+ | } | ||
+ | #sidemenu ul ul li:hover > a, | ||
+ | #sidemenu ul ul li.open > a, | ||
+ | #sidemenu ul ul li.active > a { | ||
+ | background: #424852; | ||
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+ | } | ||
+ | #sidemenu ul ul li:first-child > a { | ||
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+ | padding-left: 30px; | ||
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+ | #sidemenu > ul > li > ul > li:last-child > a, | ||
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+ | transform: rotate(-135deg); | ||
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+ | #sidemenu ul ul li.active > a::after, | ||
+ | #sidemenu ul ul li.open > a::after, | ||
+ | #sidemenu ul ul li > a:hover::after { | ||
+ | border-color: #ffffff; | ||
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+ | |||
+ | #yolo { | ||
width:100%; | width:100%; | ||
− | + | background-color: #89e1ff; | |
− | + | position:relative; | |
− | + | z-index:-1; | |
− | + | top:-220px; | |
} | } | ||
+ | |||
#contentbox{ | #contentbox{ | ||
− | width: | + | width: 70%; |
+ | float:left; | ||
+ | left:240px; | ||
+ | position:relative; | ||
max-width: 1900px; | max-width: 1900px; | ||
min-width: 1000px; | min-width: 1000px; | ||
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margin:auto; | margin:auto; | ||
font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif; | font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif; | ||
+ | font-size:18px; | ||
} | } | ||
#cover { | #cover { | ||
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margin:auto; | margin:auto; | ||
padding:0px; | padding:0px; | ||
− | background-image: url("https://static.igem.org/mediawiki/2015/ | + | background-image: url("https://static.igem.org/mediawiki/2015/e/ed/Notebookumd.jpeg"); |
background-size: 100% ; | background-size: 100% ; | ||
background-repeat: no-repeat; | background-repeat: no-repeat; | ||
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font-size:xx-large; | font-size:xx-large; | ||
top:-200px; | top:-200px; | ||
+ | z-index:3; | ||
} | } | ||
− | |||
#bar{ | #bar{ | ||
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height:200px; | height:200px; | ||
position:absolute; | position:absolute; | ||
− | + | bottom:0px; | |
text-shadow: 4px 4px 4px white; | text-shadow: 4px 4px 4px white; | ||
+ | z-index:3; | ||
} | } | ||
+ | |||
ul.a {list-style-type: circle;font-size:18px;color:black} | ul.a {list-style-type: circle;font-size:18px;color:black} | ||
</style> | </style> | ||
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<div id='bar'> | <div id='bar'> | ||
<p style="font-size:64px"><b>Protocols</b> | <p style="font-size:64px"><b>Protocols</b> | ||
+ | </div> | ||
</div> | </div> | ||
+ | |||
+ | <div id='yolo'> | ||
+ | <div id='sidemenu'> | ||
+ | <ul> | ||
+ | |||
+ | <li><a href="#Miniprep" class="smoothScroll"><span>Miniprep</span></a></li> | ||
+ | |||
+ | <li><a href="#Ligation" class="smoothScroll"><span>Ligation</span></a></li> | ||
+ | |||
+ | <li><a href="#Transformation" class="smoothScroll"><span>Transformation</span></a></li> | ||
+ | |||
+ | <li><a href="#3Assembly" class="smoothScroll"><span>3A Assembly</span></a></li> | ||
+ | |||
+ | <li><a href="#REDigest" class="smoothScroll"><span>RE Digest</span></a></li> | ||
+ | |||
+ | <li><a href="#Gibsonassembly" class="smoothScroll"><span>Gibson Assembly</span></a></li> | ||
+ | |||
+ | <li><a href="#PCR" class="smoothScroll"><span>PCR</span></a></li> | ||
+ | |||
+ | <li><a href="#Gelextract" class="smoothScroll"><span>Gel Extract</span></a></li> | ||
+ | <li><a href="#FPLC" class="smoothScroll"><span>FPLC</span></a></li> | ||
+ | </ul> | ||
</div> | </div> | ||
− | + | ||
<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="Miniprep"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>Miniprep</b></a> | <b>Miniprep</b></a> | ||
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<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="Ligation"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>Ligation</b></a> | <b>Ligation</b></a> | ||
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<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="Transformation"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>Transformation</b></a> | <b>Transformation</b></a> | ||
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<li>- Heat shock at 42°C fro 30 seconds</li> | <li>- Heat shock at 42°C fro 30 seconds</li> | ||
<li>- Add 1 mL SOC media to the cells</li> | <li>- Add 1 mL SOC media to the cells</li> | ||
− | <li>- Incubate for 60 minutes at | + | <li>- Incubate for 60 minutes at 37°C</li> |
<li>- Plate 200 µL </li> | <li>- Plate 200 µL </li> | ||
<li>- Incubate at 37°C</li> | <li>- Incubate at 37°C</li> | ||
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</div> | </div> | ||
<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="3Assembly"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>3A Assembly</b></a> | <b>3A Assembly</b></a> | ||
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<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="REDigest"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>RE Digest</b></a> | <b>RE Digest</b></a> | ||
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<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="Gibsonassembly"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
− | <b> | + | <b>Gibson Assembly</b></a> |
<p style="font-size:24px;text-align:left;text-decoration: underline;"> | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
Materials: | Materials: | ||
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<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="PCR"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>PCR</b></a> | <b>PCR</b></a> | ||
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</div> | </div> | ||
<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="Gelextract"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>Gel Extract</b></a> | <b>Gel Extract</b></a> | ||
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Procedure: | Procedure: | ||
<ul class="a"> | <ul class="a"> | ||
− | <li>- | + | <li>- Cut out gel portions with scalpel</li> |
− | <li>- | + | <li>- Weigh gel slice in colorless tube. Add 3 volumes of QG buffer to 1 volume of gel</li> |
<li>- Incubate at 50 for 10 mins until gels slice dissolves. Mix by vortexing tube every 2-3 mins during incubation to help the gel dissolve | <li>- Incubate at 50 for 10 mins until gels slice dissolves. Mix by vortexing tube every 2-3 mins during incubation to help the gel dissolve | ||
</li> | </li> | ||
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<li>- Place a Q1Aquick spin column in a provided 2ml collection tube</li> | <li>- Place a Q1Aquick spin column in a provided 2ml collection tube</li> | ||
<li>- Apply sample to spin column, centrifuge for 1 min to bind dna</li> | <li>- Apply sample to spin column, centrifuge for 1 min to bind dna</li> | ||
− | <li>- | + | <li>- Discard flow through</li> |
− | <li>- | + | <li>- Wash with .75 mL buffer PE in column, centrifuge 1 minute</li> |
<li>- Place column in a clean 1.5 microcentrifuge tube</li> | <li>- Place column in a clean 1.5 microcentrifuge tube</li> | ||
<li>- 40 microliters DDH20 , let stand 1 minute, centrifuge 1 min</li> | <li>- 40 microliters DDH20 , let stand 1 minute, centrifuge 1 min</li> | ||
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</div> | </div> | ||
<div id='contentbox'> | <div id='contentbox'> | ||
+ | <a name="FPLC"> | ||
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
<b>Fast Protein Liquid Chromatography</b></a> | <b>Fast Protein Liquid Chromatography</b></a> | ||
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Procedure: | Procedure: | ||
<ul class="a"> | <ul class="a"> | ||
− | <li>- | + | <li>- Centrifuge cell lysate (balance with equal mass of water) for 60 minutes at 25000 rpm</li> |
<li>- Wash FPLC column with equilibration column (X2) with syringe</li> | <li>- Wash FPLC column with equilibration column (X2) with syringe</li> | ||
<li>- Wash out FLPC system with equilibration buffer</li> | <li>- Wash out FLPC system with equilibration buffer</li> | ||
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<li>- Wash off excess cobalt with syringe</li> | <li>- Wash off excess cobalt with syringe</li> | ||
<li>- Connect affinity column to FPLC system</li> | <li>- Connect affinity column to FPLC system</li> | ||
− | <li>- Inject | + | <li>- Inject supernatant</li> |
<li>- Wash FPLC column with equilibration column (X2) with syringe</li> | <li>- Wash FPLC column with equilibration column (X2) with syringe</li> | ||
</ul> | </ul> |
Latest revision as of 02:32, 19 September 2015