User:David K. Barclay/Notebook/Controlling Pancreas Cell Fate Using Transcription Factors/2014/03/31: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2014/03/31 Entry for User:David_K._Barclay/Notebook/Controlling_Pancreas_Cell_Fate_Using_Transcription_Factors) |
|||
Line 6: | Line 6: | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==TC1-Alpha cells transfection redo== | ||
* | *TC1-Alpha cells transfected 10 days ago were dead due to overgrowth. Transfection needs to be redone. | ||
*Two T-75 flasks(10mL at 100% confluence) of the TC1-Alpha Cells | |||
===Transfection (6 well)=== | |||
*T-75(1) container used | |||
*Each well done 1/10, 1mL detached cells to 3mL DMEM. | |||
*6 wells= 6/10 of one T-75 used. | |||
===Splitting Cells=== | |||
*Rest of T75(1) container used. | |||
*Split cells 1/5, 2mL detached cells to 8mL DMEM. | |||
*Rest of T75(1) container disposed | |||
*Protocol used for these two functions was the Splitting Cell protocol | |||
===Freezing Cells=== | |||
*Concentration: 90% FBS, 10% DMSO | |||
*T75(2) flask used, detach cells according to Splitting Cell protocol | |||
*Transfer cells to spin column and spin down | |||
*Aspirate fluid, leaving enough to cover top of pellet, re-suspend pellet | |||
*Add 5mL Freezing Serum | |||
*Remix cells in Freezing serum | |||
*1mL into tubes, 5 tubes created for freezing | |||
Revision as of 09:40, 1 April 2014
Cell Fate Switch by Synthetic Transcription Factors | <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> |
TC1-Alpha cells transfection redo
Transfection (6 well)
Splitting Cells
Freezing Cells
|