User:Karmella Haynes/Notebook/ASU courses/2012/10/24

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10/24/12

  • Check results ofr section 1
  • Prep for week 2 (10/24/12, section 1): real DNA PCR, kits and master mixes



Check results (spot check)

  • Run 5 μL (1/20 sample) on a 1% gel (etBr)
  • Marker: [1]
Group 1
1. ctrl
2. ctrl
3. plasmid
4. plasmid
5. plasmid
6. none
7. none
8. none

Group 2
1. ctrl
2. ctrl
3. none
4. none
5. none
6. plasmid
7. plasmid
8. plasmid



Prep for Week 2, Wednesday (Sections 2, 3)

  • Assign biomarker status to fictional patients on Patient Assignment Table (Google doc)
  • Put into baggies: 1 marker, 1 PCR tube rack, 1 trimmed plastic bulb pipette (to be fitted into disposable sterile pipette tips, provided by Garcia)
  • Master Mixes
    • DONE: Promega colorless master mix: 8x 50 μL aliquots/ lab group (Joseph Mejewski) in PCR strip tubes; 48 total
    • To do: DNA/ primer mixes: 1x negative, 1x positive, 3x patient 1, 3x patient 2 (8 samples total/ lab group)

DNA/primer mixes: Make ~1/3 "patients" positive for marker

Section Groups Total Patients Pos ctrls Pos patients x3 reps TOTAL (+)'s Neg ctrls Neg patients x3 reps TOTAL (-)'s
Thurs 9:30 am 17 34 17 36 53 17 66 83
Thurs 1:30 pm 17 34 17 36 53 17 66 83


Per Section!

Reagent (+) Mix+2 (x55) (-) Mix+2 (x85)
Primer 1 (P0001) 1.0 55.0 1.0 85.0
Primer 2 (P0002) 1.0 55.0 1.0 85.0
Kozak plasmid (54 ng/μL) 0.2 11 --- ---
dH2O 47.8 2629.0 48.0 4080
  50.0 2750 50.0 4250


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