Team:Newcastle/16 June 2010

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(New page: =PCR purification protocol= # Add 5 volumes of Buffer PBI to 1 volume of the PCR reaction and mix. If the color of the mixture is orange or violet, add 10 µl of 3 M sodium acetate, pH 5.0...)
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Revision as of 08:55, 17 June 2010

PCR purification protocol

  1. Add 5 volumes of Buffer PBI to 1 volume of the PCR reaction and mix. If the color of the mixture is orange or violet, add 10 µl of 3 M sodium acetate, pH 5.0, and mix. The color of the mixture will turn yellow.
  2. Place a QIAquick column in a 2 ml collection tube.
  3. To bind DNA, apply the sample to the QIAquick column and centrifuge for 30-60s. Discard flow-through and place the column back into the same tube.
  4. To wash, add 0.75 ml Buffer PE to the QIAquick column and centrifuge for 30-60s. Discard flow-through and place the column back into the same tube.
  5. Centrifuge the column in a 2 ml collection tube for 1 min.
  6. Place each column in a clean 1.5 ml microcentrifuge tube.
  7. To elute DNA, add 50 µl water ti the center of the QIAquick membrame and centrifuge the column for 1 min. For increased DNA concentration, add 30 ml elution buffer to the center of the QIAquick membrane, let the column st
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