User:Karmella Haynes/Notebook/ASU courses/2014/03/22: Difference between revisions

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(Autocreate 2014/03/22 Entry for User:Karmella_Haynes/Notebook/ASU_courses)
 
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==Date==
==03/22/14==
* To-do list
* 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)
|}
 


----
----


Content
'''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'''<br>
* 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
 
{| 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 || 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 || --- || ---
|-
| dH<sub>2</sub>O || 47.8 || 2676.8 || 48.0 || 2688.0
|-
| &nbsp;|| 50.0 || 2800 || 50.0 || 2800
|}
 
 
 
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__NOTOC__
[[category:OWWLabNotebookV1]]
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Revision as of 16:26, 21 March 2014

ASU Courses <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>

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


|}

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