Team:UT Dallas/Protocols

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Protocols

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

Gel Purification Protocol (from QIAquick Gel Extraction Kit)

Gel Electrophoresis 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

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UT Dallas

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Notebook

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            <h2><span> Protocols</span></h2>
-           <p> <font size="4" face="arial" color = "#10314B">Ligation Protocol</font></p>
+           <p> <font size="5" face="arial" color = "#10314B">Ligation Protocol</font></p>
-           <p> <font size="4" face="arial" color = "#10314B">Gel Purification Protocol (from QIAquick Gel Extraction Kit)</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
-           <p> <font size="4" face="arial"color = "#10314B">Gel Electrophoresis Protocol</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Preparing LB+Appropriate Antibiotic Protocol</font></p>
+           <p> <font size="5" face="arial" color = "#10314B">Gel Purification Protocol (from QIAquick Gel Extraction Kit)</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Gel Electrophoresis Protocol</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Preparing Competent Cells Protocol</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Preparing LB+Appropriate Antibiotic Protocol</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Miniprep Protocol (from QIAprep Spin Miniprep Kit)</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Preparing Glycerol Stock Protocol</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Preparing Competent Cells Protocol</font></p>
-           <p> <font size="4" face="arial"color = "#10314B">Transformation Protocol</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Miniprep Protocol (from QIAprep Spin Miniprep Kit)</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Preparing Glycerol Stock Protocol</font></p>
+           <p> <font size="5" face="arial"color = "#10314B">Transformation Protocol</font></p>
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