Daniel:Notebook/ComboLock/2017-2-2

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Revision as of 17:43, 2 February 2017 by >Djacobse (Created page with "=RCA Time Test (Started Friday Jan 27)= Back to Calendar ==Phosphorothiorate RCA Primer (Started [[Dani...")
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RCA Time Test (Started Friday Jan 27)

Back to Calendar

Phosphorothiorate RCA Primer (Started Yesterday)

  1. qPCR
    1. Make qPCR master mix according to following recipe
      1. 361.2 uL nfH2O
      2. 17.2 uL AmpF6.4
      3. 430 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. 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 280 uL TBE, 70 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

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