03/18/13
- Supply list
- Prep for week 2 (03/19/13): 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 dye (Dr. Garcia)
|
| Staples reclosable poly bags, 2 mil, clear | Staples | 427331 | 1000/pk | 1
|
Prep for Week 2, Tuesday
- Assign biomarker status to fictional patients on Patient Assignment Table
- Two patients per team
- Master Mixes
- Promega colorless master mix: 8x 50 μL aliquots/ lab group (Joseph Mejewski) in PCR strip tubes; 48 total
- DNA/ primer mixes: 1x negative, 1x positive, 3x patient 1, 3x patient 2 (8 samples total/ lab group)
- Put into each rack: 1 set of DNA/ primer mix (labeled +, -, 1, 1, 1, 2, 2, 2), and 2 sets of 4 Promega mix
- Label each rack with patient ID's (followed by 1 or 2)
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
|
| Tues 9:00 | 10 | 20 | 10 | 24 | 34 | 10 | 36 | 46
|
| Reagent | (+) | Mix+2 (x36) | (-) | Mix+2 (x48)
|
| Primer 1 (P0001) | 1.0 | 36.0 | 1.0 | 48.0
|
| Primer 2 (P0002) | 1.0 | 36.0 | 1.0 | 48.0
|
| Kozak plasmid (54 ng/μL) | 0.2 | 7.2 | --- | ---
|
| dH2O | 47.8 | 1720.8 | 48.0 | 2304
|
| | 50.0 | 1800 | 50.0 | 2400
|
|
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