Daniel:Notebook/ComboLock/2017-6-16: Difference between revisions

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[[Daniel:Notebook/ComboLock|Back to Calendar]]
[[Daniel:Notebook/ComboLock|Back to Calendar]]


==Protocol-Part 5==
==Protocol-Part 5 (From [[Daniel:Notebook/ComboLock/2017-6-15|Yesterday]])==


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Revision as of 18:53, 16 June 2017

Antibody Detection Test-Positive Control (Started Wednesday)

Back to Calendar

Protocol-Part 5 (From Yesterday)

  1. Bglii digestion
    1. Make the following reactions; Do not add Bglii yet
    2. Reagent Single Rxn Vol Master Mix (6.2X)
      RCA Rxn 7 0
      10X Buffer 3.1 2 12.4
      Bglii 1 6.2
      nfH2O 10 62
      Total 20 80.6
    3. Aliquot 12 uL master mix into new tube
    4. Add 7 uL appropriate sample
    5. Mix with vortexer, spin down, and add 1 uL Bglii
    6. Incubate 2 hr 37C
    7. Heat kill with 65C for 20 min
    8. Hold at 12C
  • qPCR
    1. Make the following 7.2X master mix (1X)
      1. 7.2 uL 10 uM AmpF6.4Sol
      2. 151.2 uL (21) nfH2O
      3. 180 uL 2X Kapa SYBR Fast Master Mix
    2. Add 47 uL master mix to each lane
    3. Add 1 uL AmpR and 2 uL sample according to plate layout
    4. Use the following protocol