User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/12: Difference between revisions
From OpenWetWare
(fix raw html notebook nav) |
|||
(3 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"| | |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]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 24: | Line 24: | ||
# 0.5 | # 0.5 | ||
# 1.0 | # 1.0 | ||
# 2.0 | # 2.0 (induced on 6/02/13, 10 days) | ||
* Use 1:200 dilution of rabbit (poly) anti-luc | * Use 1:200 dilution of rabbit (poly) anti-luc (in 15 μL Gentex horse serum blocking solution per well) | ||
* Incubate under parafilm "covers" overnight at 4°C | * Incubate under parafilm "covers" overnight at 4°C in box with wet paper towels | ||
Line 59: | Line 59: | ||
> Add 7.5 μL Lipofectamine LTX to each DNA+Opti-MEM+PLUS reagent sample. 30 min/ room temp <br> | > Add 7.5 μL Lipofectamine LTX to each DNA+Opti-MEM+PLUS reagent sample. 30 min/ room temp <br> | ||
> Add 600 μL complexes to each well (final volume of medium = 4 ml/well)<br> | > Add 600 μL complexes to each well (final volume of medium = 4 ml/well)<br> | ||
> Grow cells at 37°C overnight<br> | > Grow cells at 37°C overnight<br> | ||
Latest revision as of 22:45, 26 September 2017
Pc-TF Genomics | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||||||||||||||||
06/12/13
Gal4-EED/luc cells IFC
Lipo transfection
> Add 570 μL Opti-MEM to each 20 μL DNA sample
|