User:Karmella Haynes/Notebook/PcTF Genomics/2014/11/15: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2014/11/15 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
(fix raw html notebook nav)
 
(2 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
Line 8: Line 8:




==mm/dd/yy==
==11/15/14==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
* Transfection: KAH126/MV2 into U2OS
 


----
----
'''Line item 1'''<br>
'''Transfection: KAH126/MV2 into U2OS'''<br>
> Samples
Strategy: instead of trying to increase transfection efficiency, do a "quasi-subcloning" where transfected cells are separated into smaller wells, and wells showing highest RFP will be selected, pooled, and re-plated.
 
* 6-well plate seeded on 11/14/14 (~2.5E5 cells/well)
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
* Change medium to p/s-free before transfection (4 mL per well)
|-valign="top"
* Use standard Lipofectamine LTX procedure w/ 2000 ng plasmid
| <u>Reagent</u> || <u>Volume</u>
|-
| reagent 1 || # μL
|-
| reagent 2 || #
|-
| reagent 3 || #
|-
| reagent 4 || #
|-
| dH<sub>2</sub>O || #
|-
| &nbsp; || # μL
|}


--> Reaction conditions
# KAH126/MV2
# mock
# no transfection


* Grow overnight, look for RFP next day
* Next day: harvest transfected cells and use 1/2 (5.0E5) for 2 different plates
** 24-well plate - ~2.1E4 cells/well
** 96-well plate - ~5.2E4 cells/well




Latest revision as of 00:31, 27 September 2017

Pc-TF Genomics Main project page
Previous entry      Next entry


11/15/14

  • Transfection: KAH126/MV2 into U2OS


Transfection: KAH126/MV2 into U2OS
Strategy: instead of trying to increase transfection efficiency, do a "quasi-subcloning" where transfected cells are separated into smaller wells, and wells showing highest RFP will be selected, pooled, and re-plated.

  • 6-well plate seeded on 11/14/14 (~2.5E5 cells/well)
  • Change medium to p/s-free before transfection (4 mL per well)
  • Use standard Lipofectamine LTX procedure w/ 2000 ng plasmid
  1. KAH126/MV2
  2. mock
  3. no transfection
  • Grow overnight, look for RFP next day
  • Next day: harvest transfected cells and use 1/2 (5.0E5) for 2 different plates
    • 24-well plate - ~2.1E4 cells/well
    • 96-well plate - ~5.2E4 cells/well



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2014/11/15&oldid=1015845"