User:Karmella Haynes/Notebook/ASU courses/2013/10/28: Difference between revisions

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===03/18/13==
==03/18/13==
* Supply list
* Supply list
* Prep for week 2 (03/19/13): PCR, kits and master mixes
* Prep for week 2 (10/30/13): PCR, kits and master mixes




Line 30: Line 30:
|-
|-
| SYBR green dye (Dr. Garcia)
| SYBR green dye (Dr. Garcia)
|-
| Staples reclosable poly bags, 2 mil, clear || Staples || 427331 || 1000/pk || 1
|}
|}


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----
----


'''Prep for Week 2, Tuesday'''
'''Prep for Week 2'''


* Assign biomarker status to fictional patients on Patient Assignment Table
* Assign biomarker status to fictional patients on Patient Assignment Table
* Two patients per team
* Two patients per team
* 36 total teams (18 per section on Wednesday AM/PM)
* Assisted by Alfonso Rivera and Zazu Maloi


* '''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 in PCR strip tubes; make 45 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)


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| 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'''
|-
|-
| Tues 9:00 || 10     || 20                || 10          || 24                          || '''34'''               || 10          || 36                             || '''46'''
| Wed AM || 18    || 36      || 18     || 18x3=54      || '''54'''  || 18      || 18x3=54    || '''54'''
|-
| Wed PM || 18    || 36       || 18      || 18x3=54      || '''54'''  || 18      || 18x3=54    || '''54'''
|}
|}




{|
{|
| Reagent || (+) || Mix+2 (x36) || (-) || Mix+2 (x48)
| Reagent || (+) || Mix+2 (x56) || (-) || Mix+2 (x56)
|-
|-
| Primer 1 (P0001) || 1.0 || 36.0 || 1.0 || 48.0
| Primer 1 (P0001) || 1.0 || 56.0 || 1.0 || 56.0
|-
|-
| Primer 2 (P0002) || 1.0 || 36.0 || 1.0 || 48.0
| Primer 2 (P0002) || 1.0 || 56.0 || 1.0 || 56.0
|-
|-
| Kozak plasmid (54 ng/μL) || 0.2 || 7.2 || --- || ---
| Kozak plasmid (54 ng/μL) || 0.2 || 11.2 || --- || ---
|-
|-
| dH<sub>2</sub>O || 47.8 || 1720.8 || 48.0 || 2304
| dH<sub>2</sub>O || 47.8 || 2676.8 || 48.0 || 2688.0
|-
|-
| &nbsp;|| 50.0 || 1800 || 50.0 || 2400
| &nbsp;|| 50.0 || 2800 || 50.0 || 2800
|}
|}


Revision as of 14:52, 28 October 2013

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03/18/13

  • 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 dye (Dr. Garcia)


Prep for Week 2

  • Assign biomarker status to fictional patients on Patient Assignment Table
  • Two patients per team
  • 36 total teams (18 per section on Wednesday AM/PM)
  • Assisted by Alfonso Rivera and Zazu Maloi
  • Master Mixes
    • Promega colorless master mix: 8x 50 μL aliquots/ lab group in PCR strip tubes; make 45 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 AM 18 36 18 18x3=54 54 18 18x3=54 54
Wed PM 18 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


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