Haynes Lab:Notebook/Short Projects/2014/10/20

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10/20/14 - Jonah Thomas

  • Reviewed the results of the transfection mini-experiment (see below for results)
  • Plated new bacteria for KAH60/pcVn transduction


Transfection Mini-Experiment: Results

Note: Plate #1 was left as a control and Plate #2 was spun in a centrifuge (this was completed in hopes of increasing plasmid uptake)

  • Plate #1 and Plate #2 both showed RFP signal
    • The RFP signal from Plate #1 was stronger than the RFP signal from Plate #2
    • It is hypothesized that the cells on Plate #2 died because of their increased sensitivity due to spinning; in the future, p/s-free medium should be used to prevent this from happening

Microscope Images:

  • Plate #1, Well 1A - Transfected with 1160ng KAH60/pcVn

Light microscope image of mammalian cells transfected with KAH60/pcVn.  This plate was the control group for the transfection mini-experiment. RFP signal of mammalian cells transfected with KAH60/pcVn.  This plate was the control group for the transfection mini-experiment.

Light microscope image of mammalian cells transfected with KAH60/pcVn.  This plate was the control group for the transfection mini-experiment. RFP signal of mammalian cells transfected with KAH60/pcVn.  This plate was the control group for the transfection mini-experiment.

  • Plate #1, Well 1B - Mock transfection with DH2O

Light microscope image of mammalian cells that were mock transfected with DH2O.  This plate was the control group for the transfection mini-experiment. RFP signal of mammalian cells that were mock transfected with DH2O.  This plate was the control group for the transfection mini-experiment.

  • Plate #2, Well 2A - Transfected with 1160ng KAH60/pcVn

Light microscope image of mammalian cells transfected with KAH60/pcVn.  This plate was centrifuged to increase plasmid uptake during transfection. RFP signal of mammalian cells transfected with KAH60/pcVn.  This plate was centrifuged to increase plasmid uptake during transfection.

Light microscope image of mammalian cells transfected with KAH60/pcVn.  This plate was centrifuged to increase plasmid uptake during transfection. RFP signal of mammalian cells transfected with KAH60/pcVn.  This plate was centrifuged to increase plasmid uptake during transfection.

  • Plate #2, Well 2B - Mock transfection with DH2O

Light microscope image of mammalian cells that were mock transfected with DH2O.  This plate was centrifuged to increase plasmid uptake during transfection. RFP signal of mammalian cells that were mock transfected with DH2O.  This plate was centrifuged to increase plasmid uptake during transfection.

  • Because of the success of this transfection mini-experiment, a full-blown transfection will be completed with week.
  • Multiple minipreps (4 in total) will be completed in order to increase the final concentration of KAH60/pcVn plasmids


Bacteria Plated for Minipreps

  • New DH5α-Turbo cells were plated and incubated for KAH60/pcVn transduction



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