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
|
| Wed 9:30 |
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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