User:Karmella Haynes/Notebook/ASU courses/2012/10/23: Difference between revisions

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|-
|-
| Product || Vendor || Cat # || Unit || Quantity
| Product || Vendor || Cat # || Unit || Quantity
|GoTaq colorless PCR Master Mix || Promega || M7133 || 1000 rxns
|-
|-
| 0.2 mL strip tubes & caps || BioExpress || T-3135-2 || 120 strips
|GoTaq colorless PCR Master Mix || Promega || M7133 || 1000 rxns || 1
|-
|-
|Seal-Rite graduated microcentrifuge tubes, 1.5 mL, natural, polypropylene, sterile USA Scientific 1615-5510 5x100
| 0.2 mL strip tubes & caps || BioExpress || T-3135-2 || 120 strips || 1
|-
|-
| Gene<ate 96-well PCR Tube Storage Rack BioExpress R-7909-2 5 racks
| Seal-Rite graduated microcentrifuge tubes, 1.5 mL, natural, polypropylene, sterile || USA Scientific || 1615-5510 || 5x100 || 1
|-
|-
| Transfer pipet, 1 mL, sterile Specturm Lab Products 140-98685-083 500/pk
| Genemate 96-well PCR Tube Storage Rack || BioExpress || R-7909-2 || 5 racks || 4
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| Black fine point lab markers Life Science Products 1401 10/pk
| Transfer pipet, 1 mL, sterile || Specturm Lab Products || 140-98685-083 || 500/pk || 2
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| Black fine point lab markers || Life Science Products || 1401 || 10/pk || 2
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| SYBR green dye (Dr. Garcia)
| SYBR green dye (Dr. Garcia)
|-
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| Staples reclosable poly bags, 2 mil, clear || Staples || 427331 || 1000/pk
| Staples reclosable poly bags, 2 mil, clear || Staples || 427331 || 1000/pk || 1
|}
|}


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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)


* '''Master Mixes'''<br>
* '''Master Mixes'''<br>

Revision as of 19:06, 24 October 2012

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Date

  • Supply list
  • Prep for week 2: real DNA PCR
  • 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
Transfer pipet, 1 mL, sterile Specturm Lab Products 140-98685-083 500/pk 2
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, Wednesday (Section 1)

  • 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)
  • 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)

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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