User:Karmella Haynes/Notebook/ASU courses/2012/10/24: Difference between revisions

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==Date==
==10/24/12==
* To-do list
* 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]]
|}
 
 


----
----


Content
'''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
|-
| &nbsp;|| 50.0 || 2750 || 50.0 || 4250
|}
 


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Revision as of 16:16, 2 January 2013

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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: [1]
Group 1
1. ctrl
2. ctrl
3. plasmid
4. plasmid
5. plasmid
6. none
7. none
8. none

Group 2
1. ctrl
2. ctrl
3. none
4. none
5. none
6. plasmid
7. plasmid
8. plasmid


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
    • 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

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!

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 --- ---
dH2O 47.8 2629.0 48.0 4080
  50.0 2750 50.0 4250


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