Team:UT Dallas/Protocols

From 2011.igem.org

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           <h2><span> Protocols</span></h2>
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           <h3><span>Protocols</span></h3>
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  <blockquote><h2><span></span>Ligation Protocol</h2>
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  <h3><span></span>Gel Purification Protocol (from QIAquick Gel Extraction Kit)</h3>
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  <h3><span></span>Gel Electrophoresis Protocol</h3>
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  <h3><span></span>Digestion Protocol</h3>
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  <h3><span></span>Preparing LB + Appropriate Antibiotic Protocol</h3>
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  <h3><span></span>Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)</h3>
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  <h3><span></span>Preparing Competent Cells Protocol</h3>
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  <h3><span></span>Miniprep Protocol (from QIAprep Spin Miniprep Kit)</h3>
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  <h3><span></span>Preparing Glycerol Stock Protocol</h3>
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<h3><span></span>Transformation Protocol</h3>
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           <p> <font size="5" face="arial" color = "#10314B">Ligation Protocol</font></p>
           <p> <font size="5" face="arial" color = "#10314B">Ligation Protocol</font></p>
           <li type = "square">Determine insert to vector ratios<li type = "square">Calculate the amount of insert needed if 50ng of vector is used (can use different amount of vector)<li type = "square">In a PCR tube add the following:<blockquote><li type = "circle">50ng of vector<li type = "circle">Amount of insert based on ratios (calculated in second step)<li type = "circle">2uL of buffer<li type = "circle">2uL of DNA ligase<li type = "circle">Amount of water to bring total volume to 20uL</blockquote><li type = "square">Incubate overnight at 14oC<p>Note: We used T4 DNA ligase and buffer from NEB
           <li type = "square">Determine insert to vector ratios<li type = "square">Calculate the amount of insert needed if 50ng of vector is used (can use different amount of vector)<li type = "square">In a PCR tube add the following:<blockquote><li type = "circle">50ng of vector<li type = "circle">Amount of insert based on ratios (calculated in second step)<li type = "circle">2uL of buffer<li type = "circle">2uL of DNA ligase<li type = "circle">Amount of water to bring total volume to 20uL</blockquote><li type = "square">Incubate overnight at 14oC<p>Note: We used T4 DNA ligase and buffer from NEB
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          <p> <font size="5" face="arial" color = "#10314B">Gel Purification Protocol (from QIAquick Gel Extraction Kit)</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Gel Electrophoresis Protocol</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Preparing LB+Appropriate Antibiotic Protocol</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Preparing Competent Cells Protocol</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Miniprep Protocol (from QIAprep Spin Miniprep Kit)</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Preparing Glycerol Stock Protocol</font></p>
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          <p> <font size="5" face="arial"color = "#10314B">Transformation Protocol</font></p>
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Revision as of 15:39, 17 August 2011

biz solution

Protocols

Ligation Protocol

Gel Purification Protocol (from QIAquick Gel Extraction Kit)

Gel Electrophoresis Protocol

Digestion Protocol

Preparing LB + Appropriate Antibiotic Protocol

Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)

Preparing Competent Cells Protocol

Miniprep Protocol (from QIAprep Spin Miniprep Kit)

Preparing Glycerol Stock Protocol

Transformation Protocol

Ligation Protocol

  • Determine insert to vector ratios
  • Calculate the amount of insert needed if 50ng of vector is used (can use different amount of vector)
  • In a PCR tube add the following:
  • 50ng of vector
  • Amount of insert based on ratios (calculated in second step)
  • 2uL of buffer
  • 2uL of DNA ligase
  • Amount of water to bring total volume to 20uL
  • Incubate overnight at 14oC

    Note: We used T4 DNA ligase and buffer from NEB

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