User:Karmella Haynes/Notebook/ASU courses/2013/03/18

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(03/18/13)
 
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* Assign biomarker status to fictional patients on Patient Assignment Table
* Assign biomarker status to fictional patients on Patient Assignment Table
-
* 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)
+
* Two patients per team
* '''Master Mixes'''<br>
* '''Master Mixes'''<br>
** Promega colorless master mix: 8x 50 μL aliquots/ lab group (Joseph Mejewski) in PCR strip tubes; 48 total  
** 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)
** 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
DNA/primer mixes: Make ~1/3 "patients" positive for marker
Line 52: Line 55:
| Section || Groups || Total Patients || Pos ctrls || Pos patients x3 reps || '''TOTAL (+)'s''' || Neg ctrls || Neg patients x3 reps || '''TOTAL (-)'s'''
| 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'''
+
| Tues 9:00 || 10      || 20                || 10          || 24                          || '''34'''                || 10          || 36                              || '''46'''
|}
|}

Current revision

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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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