Difference between revisions of "Team:UMaryland/Notebook2"

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<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
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<b>Ligation</b></a>
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<p style="font-size:24px;text-align:left;text-decoration: underline;">
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Materials:
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<ul class="a">
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  <li>- 2 µL PSBIA3 digest</li>
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  <li>- 2 µL upstream digest (pBAD/ sRNBC)</li>
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  <li>- 2 µLdownstream digest (miraculin / const_GFP)</li>
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  <li>- 1 µL T4 DNA ligase</li>
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  <li>- 2 µL T4 DNA ligase 10x rxn buffer</li>
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</ul>
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<p style="font-size:24px;text-align:left;text-decoration: underline;">
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Procedure:
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<ul class="a">
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  <li>- Combine reagents in a clean PCR tube</li>
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  <li>- Let stand 10 mins at room temperature/ no heat kill</li>
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</ul>
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<br>
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<br>
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</div>
  
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<div id='contentbox'>
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<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
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<b>Transformation</b></a>
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<p style="font-size:24px;text-align:left;text-decoration: underline;">
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Materials:
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<ul class="a">
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  <li>- 50 µL cells (DH5alpha)</li>
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  <li>- 2 µL DNA </li>
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</ul>
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<p style="font-size:24px;text-align:left;text-decoration: underline;">
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Procedure:
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<ul class="a">
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  <li>- Add DNA to cells</li>
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  <li>- Incubate on ice for 30 minutes</li>
 +
  <li>- Heat shock at 42° fro 30 seconds</li>
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  <li>- Add 1 mL SOC media to the cells</li>
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  <li>- Incubate for 60 minutes at 37°</li>
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  <li>- Plate 200 µL </li>
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  <li>- Incubate at 37°</li>
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</ul>
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<br>
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<br>
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</div>
 
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Revision as of 10:13, 18 September 2015