Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2014/12/16: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
| (One intermediate revision by the same user not shown) | |||
| Line 23: | Line 23: | ||
==Notes== | ==Notes== | ||
To make the transfection solution, in a 1.5mL microcentrifuge tube add: | |||
* 20μL DNA in H<sub>2</sub>O | |||
* 570μL Opti-MEM solution | |||
* 2.5μL PLUS reagent | |||
* wait 5 minutes at room temp. | |||
* 7.5μL Lipofectamine LTX | |||
* wait 30 minutes at room temp. | |||
* begin transfection<br><br> | |||
==Results/Conclusions== | ==Results/Conclusions== | ||
Three wells were treated with DNA, one was treated with water (negative control). Well plate is in the upper incubator, middle shelf.<br><br> | |||
*'''[[User:David Benjamin Nyer|David Benjamin Nyer]] | *'''[[User:David Benjamin Nyer|David Benjamin Nyer]] 18:36, 16 December 2014 (EST)''': | ||
<!-- ##### 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. ##### --> | ||
__NOTOC__ | __NOTOC__ | ||
Revision as of 16:36, 16 December 2014
 RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines
|
<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> |
SummaryIn order to introduce a new gene into the mammalian cancer cells, we need to make the cells competent and present them with a prepared plasmid containing our gene of interest along with a selectable marker. Lipofectamine is a tranfection reagent that produces positively charged liposomes that envelop the plasmids to be introduced to the cells and encourage passage through the negatively charged cell membrane into the cells. Goals
Protocolmammalian cell transfection with liopfectamine Materials
NotesTo make the transfection solution, in a 1.5mL microcentrifuge tube add:
Results/ConclusionsThree wells were treated with DNA, one was treated with water (negative control). Well plate is in the upper incubator, middle shelf.
|
|
