Team:Calgary/Notebook/Protocols/Process10

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TITLE=cDNA Synthesis/Reverse Transcription Reaction|
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TITLE=PCR Purification|
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  <h2 style="color:#0066CC">PCR Purification</h2>
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  <p>The kit we used over the summer was purchased from Qiagen. This kit is
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<p>The kit we used over the summer was purchased from Qiagen. This kit is
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used to purify PCR products, as well DNA products in enzymatic reactions, particularly, restriction digests.
used to purify PCR products, as well DNA products in enzymatic reactions, particularly, restriction digests.
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Latest revision as of 03:21, 29 September 2011


PCR Purification

The kit we used over the summer was purchased from Qiagen. This kit is used to purify PCR products, as well DNA products in enzymatic reactions, particularly, restriction digests.

  1. Add 5 volumes of Buffer PB to 1 volume of PCR product and then mix.
  2. Place a spin column in a 2 mL collection tube.
  3. Apply the mixture of PB buffer + PCR product through the QIA column and centrifuge for 1 minute.
  4. Discard the flow-through and place the column back in the tube.
  5. Wash the product using 0.75 mL buffer PE and then centrifuge for 1 min.
  6. Discard the flow-through and place the column back into the tube.
    7. Centrifuge the column inside the tube to discard the additional fluid in the column.
  7. Elute the DNA into a clean microcentrifuge tube with buffer EB.








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