Team:Paris Saclay/Protocols/PCR clean-up

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(→PCR clean-up)
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# After the PCR, pool your PCR result in an eppendorf 1,5ml.
# After the PCR, pool your PCR result in an eppendorf 1,5ml.
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# Add 200µl NTI.
+
# Add double the amount of PCR result in NTI.
# Centrifuge at 11000g, 30 seconds RT.
# Centrifuge at 11000g, 30 seconds RT.
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# Discard the supernatant, wash first time with 700µl NT3.
+
# Discard the supernatant, wash a first time with 700µl NT3.
# Centrifuge at 11000g, 30 secondes RT.
# Centrifuge at 11000g, 30 secondes RT.
# Discard the supernatant, wash a second time with 700µl NT3.
# Discard the supernatant, wash a second time with 700µl NT3.
# Centrifuge at 11000g, 30 seconds RT.
# Centrifuge at 11000g, 30 seconds RT.
# Centrifuge the column empty.
# Centrifuge the column empty.
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# Add 20µl MilliQ water and centrifuge at 11000g, 1 min RT.
+
# Add 15-30µl elution buffer and centrifuge at 11000g, 1 min RT.
# Collect your DNA in an eppendorf 1,5ml.
# Collect your DNA in an eppendorf 1,5ml.
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Latest revision as of 11:53, 11 August 2014

PCR clean-up

  1. After the PCR, pool your PCR result in an eppendorf 1,5ml.
  2. Add double the amount of PCR result in NTI.
  3. Centrifuge at 11000g, 30 seconds RT.
  4. Discard the supernatant, wash a first time with 700µl NT3.
  5. Centrifuge at 11000g, 30 secondes RT.
  6. Discard the supernatant, wash a second time with 700µl NT3.
  7. Centrifuge at 11000g, 30 seconds RT.
  8. Centrifuge the column empty.
  9. Add 15-30µl elution buffer and centrifuge at 11000g, 1 min RT.
  10. Collect your DNA in an eppendorf 1,5ml.

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