Difference between revisions of "Team:RHIT/Protocol"
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<ul> | <ul> | ||
− | <li class="Title" id="QIAPREP">QIAprep Spin Miniprep Kit</li> | + | <li class="Title" id="QIAPREP" onmouseover="changecursor('QIAPREP')">QIAprep Spin Miniprep Kit</li> |
<ol id="QIAPREPSTEP" style="display:none"> | <ol id="QIAPREPSTEP" style="display:none"> | ||
<li class="steps"> Pellet 1-5 ml bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature.</li> | <li class="steps"> Pellet 1-5 ml bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature.</li> | ||
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− | <li class="Title" id="NEBuilder">NEBuilder HiFi DNA Assembly Reaction Protocol</li> | + | <li class="Title" id="NEBuilder" onmouseover="changecursor('NEBuilder')">NEBuilder HiFi DNA Assembly Reaction Protocol</li> |
<ol id="NEBuilderSTEP" style="display:none"> | <ol id="NEBuilderSTEP" style="display:none"> | ||
<li class="steps">Set up the following reaction on ice (for 2-3 fragment assembly):</li> | <li class="steps">Set up the following reaction on ice (for 2-3 fragment assembly):</li> | ||
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− | <li class="Title" id="BOILING">Boiling Prep Protocol</li> | + | <li class="Title" id="BOILING" onmouseover="changecursor('BOILING')">Boiling Prep Protocol</li> |
<ol id="BOILINGSTEP" style="display:none"> | <ol id="BOILINGSTEP" style="display:none"> | ||
<li class="steps">Spin down cells.</li> | <li class="steps">Spin down cells.</li> | ||
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− | <li class="Title" id="NEB">NEB High Efficiency Transformation Protocol</li> | + | <li class="Title" id="NEB" onmouseover="changecursor('NEB')">NEB High Efficiency Transformation Protocol</li> |
<ol id="NEBSTEP" style="display:none"> | <ol id="NEBSTEP" style="display:none"> | ||
<li class="steps">Thaw a tube of NE 5-alpha Competent E. coli cells on ice until the last ice crystals disappear. Mix gently and carefully pipette 50 µl of cells into a transformation tube on ice.</li> | <li class="steps">Thaw a tube of NE 5-alpha Competent E. coli cells on ice until the last ice crystals disappear. Mix gently and carefully pipette 50 µl of cells into a transformation tube on ice.</li> | ||
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− | <li class="Title" id="QIAquick">QIAquick Gel Extraction Kit</li> | + | <li class="Title" id="QIAquick" onmouseover="changecursor('QIAquick')">QIAquick Gel Extraction Kit</li> |
<ol id="QIAquickSTEP" style="display:none"> | <ol id="QIAquickSTEP" style="display:none"> | ||
<li class="steps">Excise the DNA fragment from the agarose gel with a clean, sharp scalpel.</li> | <li class="steps">Excise the DNA fragment from the agarose gel with a clean, sharp scalpel.</li> | ||
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− | <li class="Title" id="Ethanol">Ethanol Precipitation</li> | + | <li class="Title" id="Ethanol" onmouseover="changecursor('Ethanol')">Ethanol Precipitation</li> |
<ol id="EthanolSTEP" style="display:none"> | <ol id="EthanolSTEP" style="display:none"> | ||
<li class="steps">Estimate the volume of the DNA solution.</li> | <li class="steps">Estimate the volume of the DNA solution.</li> | ||
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− | <li class="Title" id="Yeast">Yeast Transformation (From Breeden Lab)</li> | + | <li class="Title" id="Yeast" onmouseover="changecursor('Yeast')">Yeast Transformation (From Breeden Lab)</li> |
<ol id="YeastSTEP" style="display:none"> | <ol id="YeastSTEP" style="display:none"> | ||
<li class="steps">Pick colonies and inoculate cultures in YPD and grow overnight.</li> | <li class="steps">Pick colonies and inoculate cultures in YPD and grow overnight.</li> |
Revision as of 17:46, 17 August 2015
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Protocols
- QIAprep Spin Miniprep Kit
- NEBuilder HiFi DNA Assembly Reaction Protocol
- Boiling Prep Protocol
- NEB High Efficiency Transformation Protocol
- QIAquick Gel Extraction Kit
- Ethanol Precipitation
- Yeast Transformation (From Breeden Lab)