# User:Karmella Haynes/Notebook/ASU courses/2013/10/28

(Difference between revisions)
 Revision as of 17:49, 28 October 2013 (view source) (→03/18/13)← Previous diff Current revision (17:52, 28 October 2013) (view source) (→03/18/13) (2 intermediate revisions not shown.) Line 55: 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 AM || 18    || 36      || 18      || 18x3=54      || '''54'''  || 18      || 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 (x56) || (-) || Mix+2 (x48) + | Reagent || (+) || Mix+2 (x56) || (-) || Mix+2 (x56) |- |- - | Primer 1 (P0001) || 1.0 || 56.0 || 1.0 || 48.0 + | Primer 1 (P0001) || 1.0 || 56.0 || 1.0 || 56.0 |- |- - | Primer 2 (P0002) || 1.0 || 56.0 || 1.0 || 48.0 + | Primer 2 (P0002) || 1.0 || 56.0 || 1.0 || 56.0 |- |- | Kozak plasmid (54 ng/μL) || 0.2 || 11.2 || --- || --- | Kozak plasmid (54 ng/μL) || 0.2 || 11.2 || --- || --- |- |- - | dH2O || 47.8 || 2676.8 || 48.0 || 2304 + | dH2O || 47.8 || 2676.8 || 48.0 || 2688.0 |- |- - |  || 50.0 || 2800 || 50.0 || 2400 + |  || 50.0 || 2800 || 50.0 || 2800 |} |}

## Current revision

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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