Matt:LabNotes/2014-1-29

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Revision as of 02:07, 1 February 2014 by >Mzcai (Created page with "===Does time after hybridization (and wash step) affect the number of signals?=== *Took the same wells from Matt:LabNotes/2014-1-28 and looked at them again 24 hours later...")
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Does time after hybridization (and wash step) affect the number of signals?[edit]

Result[edit]

' Initial 24hrs
Cy3-220nt 711 654
Cy3-300nt 357 311
Cy3-510nt 4449 3912
Cy3-1000nt 13295 12280
488-220nt 342 289
488-300nt 136 116
488-510nt 396 359
488-1000nt 20910 21370

File:RolonyCountAfter24Hours.JPG

  • There's a slight decrease for all except 488-1000nt
    • This does not support the hypothesis that longer time leads to an increase in signal as seen in: Matt:LabNotes/2014-1-27#Special Notes
    • Slight decrease can be due to photo bleaching or just variance

A different explanation: Lack of Oil[edit]

  • One effect I noticed is that if there isn't enough oil on the lens the signal may be hard to see

File:EffectOfOil.JPG

  • However it's hard to tell when the lack of signal is real and when it's due to lack of oil because the brightfield looks very similar

File:BF EffectOfOil.JPG