Haynes Lab:Notebook/Short Projects/2014/10/20: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes Lab - Short Projects</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes Lab - Short Projects</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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== | ==10/20/14 - Jonah Thomas== | ||
* | * Reviewed the results of the transfection mini-experiment (see below for results) | ||
* Plated new bacteria for KAH60/pcVn transduction | |||
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'''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 | |||
[[Image:1A_DNA_phase_1.jpg|200px|Light microscope image of mammalian cells transfected with KAH60/pcVn. This plate was the control group for the transfection mini-experiment.]] | |||
[[Image:1A_DNA_mCHERRY_1.jpg|200px|RFP signal of mammalian cells transfected with KAH60/pcVn. This plate was the control group for the transfection mini-experiment.]] | |||
[[Image:1A_DNA_phase_2.jpg|200px|Light microscope image of mammalian cells transfected with KAH60/pcVn. This plate was the control group for the transfection mini-experiment.]] [[Image:1A_DNA_mCHERRY_2.jpg|200px|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 | |||
[[Image:1B_mock_phase_1.jpg|200px|Light microscope image of mammalian cells that were mock transfected with DH2O. This plate was the control group for the transfection mini-experiment.]] | |||
[[Image:1B_mock_mCHERRY_1.jpg|200px|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 | |||
[[Image:2A_DNA_phase_1.jpg|200px|Light microscope image of mammalian cells transfected with KAH60/pcVn. This plate was centrifuged to increase plasmid uptake during transfection.]] | |||
[[Image:2A_DNA_mCHERRY_1.jpg|200px|RFP signal of mammalian cells transfected with KAH60/pcVn. This plate was centrifuged to increase plasmid uptake during transfection.]] | |||
[[Image:2A_DNA_phase_2.jpg|200px|Light microscope image of mammalian cells transfected with KAH60/pcVn. This plate was centrifuged to increase plasmid uptake during transfection.]] [[Image:2A_DNA_mCHERRY_2.jpg|200px|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 | |||
[[Image:2B_mock_phase_1.jpg|200px|Light microscope image of mammalian cells that were mock transfected with DH2O. This plate was centrifuged to increase plasmid uptake during transfection.]] | |||
[[Image:2B_mock_mCHERRY_1.jpg|200px|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 | |||
Latest revision as of 00:27, 27 September 2017
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10/20/14 - Jonah Thomas
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)
Microscope Images:
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