Daniel:Notebook/PosSequencing/2017-5-20

From ZhangLabWiki
Revision as of 20:10, 20 May 2017 by >Djacobse (Created page with "=Single Stranded Probe Binding Test (Started Yesterday)= Back to Calendar ==Protocol== <ol sta...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

Single Stranded Probe Binding Test (Started Yesterday)

Back to Calendar

Protocol

  1. Single primer qPCR
    1. Make the following master mixes
    2. Reagent uL in Single Rxn Master Mix P2 (3.2X) Master Mix BioU (3.2X) Master Mix Dual (2.2X)
      Primer 2 (10 uM) 1 3.2 3.2 2.2
      BioU Primer (10 uM) 1 0 0 2.2
      2X Kapa SYBR Master Mix 20 64 64 44
      nfH2O 17 54.4 54.4 35.2
      Total   121.6 121.6 83.6
    3. Add 37 uL or 38 uL (dual) to the appropriate lanes
    4. Add 2 uL samples and 1 (or 2) uL primer according to plate layout
    5. File:PlateLayout-20170520-SinglePrimer.png
    6. qPCR Cycles
      1. 95C 3 min
      2. 95C 3 sec
      3. 55C 30 sec
      4. 72C 20 sec
      5. plate read
      6. goto b x30
      7. 72C 2 min
      8. 16C hold