User:Karmella Haynes/Notebook/ASU courses/2012/10/24: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> ASU Courses</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> ASU Courses</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==10/24/12== | ||
* | * Check results for section 1 | ||
* Prep for week 2 (10/24/12, section 1): real DNA PCR, kits and master mixes | |||
---- | ---- | ||
'''Check results''' (spot check) | |||
* Run 5 μL (1/20 sample) on a 1% gel (etBr) | |||
* Expected size = ~200 - 300 | |||
* Marker: [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg] | |||
{| class="wikitable" width=500 | |||
|- valign="top" | |||
| Group 1<br>1. ctrl<br> 2. ctrl<br>3. plasmid<br>4. plasmid<br>5. plasmid<br>6. none<br>7. none<br>8. none<br><br>Group 2<br>1. ctrl<br> 2. ctrl<br>3. none<br>4. none<br>5. none<br>6. plasmid<br>7. plasmid<br>8. plasmid | |||
| [[Image:KAH102412_gel1.png|250px]] | |||
|} | |||
---- | |||
'''Prep for Week 2, Wednesday (Sections 2, 3)''' | |||
* Assign biomarker status to fictional patients on Patient Assignment Table (Google doc) | |||
* 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> | |||
** DONE: Promega colorless master mix: 8x 50 μL aliquots/ lab group (Joseph Mejewski) in PCR strip tubes; 48 total | |||
** To do: 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 | |||
{| {{table}} | |||
|- | |||
| Section || Groups || Total Patients || Pos ctrls || Pos patients x3 reps || '''TOTAL (+)'s''' || Neg ctrls || Neg patients x3 reps || '''TOTAL (-)'s''' | |||
|- | |||
| Thurs 9:30 am || 17 || 34 || 17 || 36 || '''53''' || 17 || 66 || '''83''' | |||
|- | |||
| Thurs 1:30 pm || 17 || 34 || 17 || 36 || '''53''' || 17 || 66 || '''83''' | |||
|} | |||
Per Section! | |||
{| {{table}} | |||
| Reagent || (+) || Mix+2 (x55) || (-) || Mix+2 (x85) | |||
|- | |||
| Primer 1 (P0001) || 1.0 || 55.0 || 1.0 || 85.0 | |||
|- | |||
| Primer 2 (P0002) || 1.0 || 55.0 || 1.0 || 85.0 | |||
|- | |||
| Kozak plasmid (54 ng/μL) || 0.2 || 11 || --- || --- | |||
|- | |||
| dH<sub>2</sub>O || 47.8 || 2629.0 || 48.0 || 4080 | |||
|- | |||
| || 50.0 || 2750 || 50.0 || 4250 | |||
|} | |||
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Latest revision as of 22:09, 26 September 2017
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10/24/12
Check results (spot check)
Prep for Week 2, Wednesday (Sections 2, 3)
DNA/primer mixes: Make ~1/3 "patients" positive for marker
|