User:Karmella Haynes/Notebook/ASU courses/2014/03/22

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03/22/14

  • Supply list
  • Prep for week 2 (10/30/13): PCR, kits and master mixes



Supply list

Product Vendor Cat # Unit Quantity
GoTaq colorless PCR Master Mix Promega M7133 1000 rxns 1
0.2 mL strip tubes & caps BioExpress T-3135-2 120 strips 1
Seal-Rite graduated microcentrifuge tubes, 1.5 mL, natural, polypropylene, sterile USA Scientific 1615-5510 5x100 1
Genemate 96-well PCR Tube Storage Rack BioExpress R-7909-2 5 racks 4
Black fine point lab markers Life Science Products 1401 10/pk 2
SYBR Green I (Promega, Dr. Garcia)



Prep for Week 2

  • Assign biomarker status to fictional patients on Patient Assignment Table
  • Two patients per team
  • 30 total teams (14 Wednesday section, 16 Thursday section)

Master Mixes

  • Promega colorless master mix (pre-made): 8x 50 μL aliquots
    • Make 35 strips, freeze in a rack, then after they are frozen distribute enough for each class section into two baggies
  • DNA/ primer mixes: 1x negative, 1x positive, 3x patient 1, 3x patient 2
    • Label positive "+"
    • Label negative "-"
    • Label next 3 tubes with single tape showing patient ID
    • Label next 3 tubes with single tape showing patient ID


DNA/primer mixes: Make ~1/4 "patients" positive for marker

Section Groups Total Patients Pos ctrls Pos patients x3 reps TOTAL (+)'s Neg ctrls Neg patients x3 reps TOTAL (-)'s
Wed 14 36 18 18x3=54 54 18 18x3=54 54
Thurs 16 36 18 18x3=54 54 18 18x3=54 54


Reagent (+) Mix+2 (x56) (-) Mix+2 (x56)
Primer 1 (P0001) 1.0 56.0 1.0 56.0
Primer 2 (P0002) 1.0 56.0 1.0 56.0
Kozak plasmid (54 ng/μL) 0.2 11.2 --- ---
dH2O 47.8 2676.8 48.0 2688.0
  50.0 2800 50.0 2800


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