User:Karmella Haynes/Notebook/PcTF Genomics/2013/07/05: Difference between revisions
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|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}}}}]]}} | ||
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| colspan="2"| | | colspan="2"| | ||
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| <u>Wells</u> || <u>Plasmid</u> || <u>DNA</u> || <u>Volume + dH<sub>2</sub>O</u> || <u>Opti-MEM</u> || <u>PLUS</u> || <u>Lipo</u> || | | <u>Wells</u> || <u>Plasmid</u> || <u>DNA</u> || <u>Volume + dH<sub>2</sub>O</u> || <u>Opti-MEM</u> || <u>PLUS</u> || <u>Lipo</u> || | ||
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| 1 || KAH160/MV1 || 2 μg || | | 1 || KAH160/MV1 || 2 μg || 5.8 μL +14.2 || 570 μL || 2.5 μL || 7.5 μL | ||
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| 2 || KAH160/MV1 || 2 μg || | | 2 || KAH160/MV1 || 2 μg || 5.8 μL +14.2 || " || " || " | ||
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| 3 || mock || none || 0 μL + 20.0 || " || " || " | | 3 || mock || none || 0 μL + 20.0 || " || " || " | ||
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| 4 || none || none || --- || --- || --- || --- | | 4 || none || none || --- || --- || --- || --- | ||
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> Add 570 μL Opti-MEM to each 20 μL DNA sample <br> | |||
> Add 2.5 μL PLUS reagent to each DNA+Opti-MEM sample. 5 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> | |||
> Grow cells at 37°C overnight<br> | |||
Results (7/06/13) | |||
* Success! Very high levels of RFP expression in wells 1 & 2 (microscopy), zero signal in mock and untreated (3 & 4) | |||
* Harvest cells for flow cytometry | |||
Latest revision as of 22:58, 26 September 2017
Pc-TF Genomics | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||
07/05/13
Lipo transfection
> Add 570 μL Opti-MEM to each 20 μL DNA sample Results (7/06/13)
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