Team:Calgary/Notebook/Protocols/Process10

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Revision as of 00:50, 29 September 2011


cDNA Synthesis/Reverse Transcription Reaction

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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