Daniel:Notebook/ComboLock/2017-1-25

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Revision as of 16:40, 25 January 2017 by >Djacobse (Created page with "==Protocol== <li>qPCR</li> <ol type="A"> <li>Make qPCR master mix according to following recipe</li> <ol type="a"> <li>361.2 uL nfH2O</li> <li>17.2 uL AmpF-CLv3</li> <li>17.2...")
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Protocol

  • qPCR
    1. Make qPCR master mix according to following recipe
      1. 361.2 uL nfH2O
      2. 17.2 uL AmpF-CLv3
      3. 17.2 uL AmpR-CLv3
      4. 430 uL Kapa SYBR Fast
    2. Add 48 uL master mix to each well
    3. Add 2 uL sample according to table
    4. qPCR Cycles
      1. 95C 3 min
      2. 95C 3 sec
      3. 55C 30 sec
      4. 72C 20 sec
      5. plate read
      6. goto b x24
      7. 72C 2 min
      8. 16C hold
  • TBE Gel
    1. Mix 80 uL TBE, 20 uL 6x loading dye
    2. Aliquot 10 uL per sample/ladder lane onto parafilm
    3. Add 2 uL of sample or ladder to correct drop
    4. Load 10 uL in to well
    5. Run gel for 22 minutes at 250V
    6. Open gel and stain with 3 uL SYBR Gold for 3 minutes
    7. Rinse gel and image in gel doc

    Results