Daniel:Notebook/ComboLock/2017-1-31

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RCA Time Test (Started Friday)[edit]

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Epicentre RCA 12 hour (Started Yesterday)[edit]

  1. qPCR
    1. Make 5.2X qPCR master mix according to following recipe
      1. 109.2 uL nfH2O
      2. 5.2 uL AmpF6.4
      3. 130 uL Kapa SYBR Fast
    2. Add 47 uL master mix to each well
    3. Add 2 uL sample and 1 uL AmpR indexed according to table
    4. Sample Name Sample Condition AmpR Index Lane Sample Vol (uL) 2X Kapa SYBR qPCR MM 10 uM Forward Primer 10 uM Reverse Primer H2O Total Volume (uL)
      Sample 1A Sample Epi-12hr 24 A1 2 25 1 1 21 50
      Sample 1B Sample Epi-12hr 24 A2 2 25 1 1 21 50
      Sample 2A No PO4 Epi-12hr 28 A3 2 25 1 1 21 50
      Sample 2B No PO4 Epi-12hr 28 A4 2 25 1 1 21 50
      NTC None   29 A5 0 25 1 1 23 50
    5. 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
  2. TBE Gel
    1. Mix 64 uL TBE, 16 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 23 minutes at 230V
    6. Open gel and stain with 3 uL SYBR Gold for 3 minutes
    7. Rinse gel and image in gel doc

Results[edit]