User:David Dreher/Notebook/Chromatin controlled cell pattern/2013/02/22 - Revision history

David Dreher: /* 02/23/13 */

David Dreher — Tue, 19 Mar 2013 03:53:16 GMT

02/23/13

←Older revision		Revision as of 03:53, 19 March 2013
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	* Did not detect D-luc signal. Perhaps D-luc imaging should start immediately after adding reagents. The D-luc signal has a short half-life.		* Did not detect D-luc signal. Perhaps D-luc imaging should start immediately after adding reagents. The D-luc signal has a short half-life.
	* Cells rounded up a bit after sitting in PBS. Next time use 10% FBS in PBS, or try DMEM without red dye.		* Cells rounded up a bit after sitting in PBS. Next time use 10% FBS in PBS, or try DMEM without red dye.
		+	* DAPI signal was barely visible


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	* HEK293 Gal4-EED, 1:4		* HEK293 Gal4-EED, 1:4
	* HEK293 Gal4-EED, 1:4 in 0.5 μg/mL puromycin (to select for Gal4-EED RNAi tuner)		* HEK293 Gal4-EED, 1:4 in 0.5 μg/mL puromycin (to select for Gal4-EED RNAi tuner)
		+
		+

David Dreher: /* 02/23/13 */

David Dreher — Mon, 18 Mar 2013 16:12:40 GMT

02/23/13

←Older revision		Revision as of 16:12, 18 March 2013
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	'''DAPI/ luciferase staining'''<br>		'''DAPI/ luciferase staining'''<br>
-	~~Note: David, enter formulas, procedures here~~

-	* D-luciferin stock solution: ~~???~~ in 1xPBS	+	* D-luciferin stock solution: 30mg/ml in 1xPBS, working solution is 150ug/ml (200x)
	* Hoescht: made by Dr. Haynes; 5000x in sterile dH<sub>2</sub>O		* Hoescht: made by Dr. Haynes; 5000x in sterile dH<sub>2</sub>O

Karmella Haynes: /* Entry title */

Karmella Haynes — Sat, 23 Feb 2013 22:39:17 GMT

Entry title

←Older revision		Revision as of 22:39, 23 February 2013
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	\| 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. ##### -->
-	==~~Entry title~~==	+	==02/23/13==
-	* ~~Insert content~~ here...	+	* DAPI/ luciferase staining
		+	* New cell plates
		+
		+	----
		+
		+	'''DAPI/ luciferase staining'''<br>
		+	Note: David, enter formulas, procedures here
		+
		+	* D-luciferin stock solution: ??? in 1xPBS
		+	* Hoescht: made by Dr. Haynes; 5000x in sterile dH<sub>2</sub>O
		+
		+
		+	* Diluted Hoescht and D-luc to working concentration (1x) in 1xPBS. Added 2 mL to each well...
		+	Used 3 wells:
		+	# DAPI + D-luc
		+	# DAPI only
		+	# PBS
		+	* Let cells sit at room temperature for 10 minutes in the dark to develop signal
		+
		+	Results/ Conclusions:
		+	* 2 mL medium is susceptible to drying at outer wells; disrupts cell growth
		+	* Did not detect D-luc signal. Perhaps D-luc imaging should start immediately after adding reagents. The D-luc signal has a short half-life.
		+	* Cells rounded up a bit after sitting in PBS. Next time use 10% FBS in PBS, or try DMEM without red dye.
		+
		+
		+	Note: add photos here
		+
		+	----
		+
		+	'''New cell plates'''<br>
		+	Dr. Haynes; For more practice staining (Monday)
		+	* HEK293 luc #4, 6-well plate, ~1x10<sup>5</sup> cells/ well, 4 mL growth medium
		+	* HEK293 Gal4-EED, 6-well plate, ~1x10<sup>5</sup> cells/ well, 4 mL growth medium
		+
		+	Stock cultures (T75 flasks)
		+	* HEK293 luc #4, 1:5
		+	* HEK293 Gal4-EED, 1:4
		+	* HEK293 Gal4-EED, 1:4 in 0.5 μg/mL puromycin (to select for Gal4-EED RNAi tuner)

Karmella Haynes: Autocreate 2013/02/22 Entry for User:David_Dreher/Notebook/Chromatin_controlled_cell_pattern

Karmella Haynes — Sat, 23 Feb 2013 22:25:43 GMT

Autocreate 2013/02/22 Entry for User:David_Dreher/Notebook/Chromatin_controlled_cell_pattern

New page

{|{{table}} width="800"
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>      </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|-
| colspan="2"|

==Entry title==
* Insert content here...


|}

__NOTOC__