Haynes Lab:Notebook/Short Projects/2014/09/19: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Jonah Thomas (talk | contribs) |
Jonah Thomas (talk | contribs) |
||
| Line 8: | Line 8: | ||
==09/19/14 - Jonah Thomas== | ==09/19/14 - Jonah Thomas== | ||
* Failed to remove plates containing transformed bacterial cells from incubator and thus, the cells became overgrown | * Failed to remove plates containing transformed bacterial cells from incubator and thus, the cells became overgrown | ||
* Measured the concentrations of the available plasmids | |||
* Measured the concentrations of the available | |||
* Performed a 1:4 passage of the dox- and dox+ cells in a T-75 flask | * Performed a 1:4 passage of the dox- and dox+ cells in a T-75 flask | ||
---- | ---- | ||
'''Plasmid Concentration Measurement''' | '''Plasmid Concentration Measurement''' | ||
| Line 18: | Line 19: | ||
Note: Measurements taken using Biotek machine - Take 3 plate | Note: Measurements taken using Biotek machine - Take 3 plate | ||
* Thawed 6 vials of plasmid | * Thawed 6 vials of plasmid at room temperature | ||
** Labeled each vial 1-6, respectively | ** Labeled each vial 1-6, respectively | ||
* Placed 2μL of plasmid on the Take 3 Plate according to Table 1 below: | * Placed 2μL of plasmid on the Take 3 Plate according to Table 1 below: | ||
| Line 41: | Line 42: | ||
|} | |} | ||
Note: It was calculated that '''9.102μL of the KAH60/vnPc plasmid (Vial 2)''' will be needed to reach the 2000ng mass necessary for plasmid for cell transfection | Note: It was calculated that '''9.102μL of the KAH60/vnPc plasmid (Vial 2)''' will be needed to reach the 2000ng mass necessary for plasmid for cell transfection. Additionally, there appeared to be ~45-50μL of this plasmid available | ||
---- | |||
'''Cell Passaging''' | |||
* HEK293 Gal4-EED cells (-dox and +dox) were passaged into a sterile T-75 flask | |||
* The standard cell passaging protocol was followed | |||
* A 1:4 passage was performed in 1% pen-strep medium | |||
---- | |||
'''Plan of Action: Week of 9/22/14''' | |||
* Monday (9/22): Transfer -dox and +dox cells to 6-well plates | |||
* Tuesday (9/23): Check cell growth | |||
* Wednesday (9/24): Perform cell transfections with KAH60/vnPc (Vial 2) | |||
** Be sure to all do mock transfections! | |||
** Depending on cell growth, this may be able to be completed on Tuesday | |||
* Thursday (9/25): Check cell growth | |||
* Friday (9/26): Check cell growth | |||
* On either Thurs. or Friday, the fluorescent microscope will be used to confirm a successful transfection of cells | |||
Note: Upon confirming a successful transfection, a cell count will be completed as well as a luciferase assay | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | ||
Revision as of 15:32, 19 September 2014
 Haynes Lab - Short Projects
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||
09/19/14 - Jonah Thomas
Note: Measurements taken using Biotek machine - Take 3 plate
Note: It was calculated that 9.102μL of the KAH60/vnPc plasmid (Vial 2) will be needed to reach the 2000ng mass necessary for plasmid for cell transfection. Additionally, there appeared to be ~45-50μL of this plasmid available
Note: Upon confirming a successful transfection, a cell count will be completed as well as a luciferase assay
| |||||||||||||||||||||||||||||||||||||||||
