User:Karmella Haynes/Notebook/ASU courses/2013/03/18: Difference between revisions

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==Date==
==03/18/13==
* To-do list
* 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
|}
 


----
----


Content
'''Prep for Week 2, Tuesday'''
 
* 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>
** 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
 
{| class="wikitable" border=1
|-
| 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 || --- || ---
|-
| dH<sub>2</sub>O || 47.8 || 1720.8 || 48.0 || 2304
|-
| &nbsp;|| 50.0 || 1800 || 50.0 || 2400
|}
 


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Revision as of 17:50, 18 March 2013

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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
  • 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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