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
|
|