Difference between revisions of "User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/05"

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==July 5, 2013==
 
==July 5, 2013==
 
===FCS Sample Prep===
 
===FCS Sample Prep===
 +
*Green fluorescent beads
 +
**10000x diluted, run 3x for 90s
 +
*PBS
 +
**To check for background, run 3x for 200s
 +
*DNA/MB
 +
**125/100pM, run 3x for 500s
 +
**100/80pM, run 3x for 500s
 +
**75/60pM, run 3x for 500s
 +
**50/40pM, run 3x for 500s
 +
**25/20pM, run 3x for 500s
 
===FCS Data===
 
===FCS Data===
====Notes====
+
Graphs generated using Igor Pro.<br.>
 +
<br.>
 +
[[Image:Avg_Beads.png]]<br.>
 +
10000x diluted green fluorescent beads were used to align the laser.  Sample was run three times, for 90s each.<br.>
 +
<br.>
 +
[[Image:Avg_PBS.png]]<br.>
 +
PBS run to check for background fluorescence.  Sample was run three times, 200s each to eliminate noise.<br.>
 +
[[Image:Avg_125pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_100pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_75pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_50pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_25pM.png]]<br.>
 +
All DNA/MB samples run three times, for 500s.<br.>
 +
<br.>
 +
Calibration Graph:<br.>
 +
[[Image:2013_0705_calibration.PNG]]
  
  

Revision as of 13:49, 5 July 2013

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July 5, 2013

FCS Sample Prep

  • Green fluorescent beads
    • 10000x diluted, run 3x for 90s
  • PBS
    • To check for background, run 3x for 200s
  • DNA/MB
    • 125/100pM, run 3x for 500s
    • 100/80pM, run 3x for 500s
    • 75/60pM, run 3x for 500s
    • 50/40pM, run 3x for 500s
    • 25/20pM, run 3x for 500s

FCS Data

Graphs generated using Igor Pro.<br.> <br.> Avg Beads.png<br.> 10000x diluted green fluorescent beads were used to align the laser. Sample was run three times, for 90s each.<br.> <br.> Avg PBS.png<br.> PBS run to check for background fluorescence. Sample was run three times, 200s each to eliminate noise.<br.> Avg 125pM.png<br.> <br.> Avg 100pM.png<br.> <br.> Avg 75pM.png<br.> <br.> Avg 50pM.png<br.> <br.> Avg 25pM.png<br.> All DNA/MB samples run three times, for 500s.<br.> <br.> Calibration Graph:<br.> 2013 0705 calibration.PNG