https://bridgeslab.sph.umich.edu/protocols/index.php?action=history&feed=atom&title=Lipofectamine_Transfection_of_C2C12_Cells 2026-06-01T21:59:19Z Revision history for this page on the wiki MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=Lipofectamine_Transfection_of_C2C12_Cells&diff=747&oldid=prev 2013-02-22T16:57:30Z

<p>changed confluence note</p> <table style="background-color: #fff; color: #202122;" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="en"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 16:57, 22 February 2013</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l5">Line 5:</td> <td colspan="2" class="diff-lineno">Line 5:</td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br></td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Materials==</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>==Materials==</div></td></tr> <tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>*Cells in a 6 well dish, plated and at <del style="font-weight: bold; text-decoration: none;">&gt;90</del>% confluence</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>*Cells in a 6 well dish, plated and at <ins style="font-weight: bold; text-decoration: none;">~70</ins>% confluence<ins style="font-weight: bold; text-decoration: none;">. Typically transfect as myocytes.</ins></div></td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Lipofectamine 2000 (Invitrogen cat# 11668-019 for 1.5mL)</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Lipofectamine 2000 (Invitrogen cat# 11668-019 for 1.5mL)</div></td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*OptiMEM</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*OptiMEM</div></td></tr> <!-- diff cache key bridgeslabproto-mw_:diff:1.41:old-746:rev-747:php=table --> </table>

Davebrid https://bridgeslab.sph.umich.edu/protocols/index.php?title=Lipofectamine_Transfection_of_C2C12_Cells&diff=746&oldid=prev 2013-02-22T16:56:06Z

<p>Wrote protocol from ethan&#039;s trials</p> <p><b>New page</b></p><div>[[Category: Cell Culture ]]<br /> [[Category: Transfection]]<br /> [[Category: Muscle Cells ]]<br /> [[Category: C2C12 Cells ]]<br /> <br /> ==Materials==<br /> *Cells in a 6 well dish, plated and at &gt;90% confluence<br /> *Lipofectamine 2000 (Invitrogen cat# 11668-019 for 1.5mL)<br /> *OptiMEM<br /> <br /> ==Required Amounts==<br /> *Calculate DNA to transfect. Start with 4 ug per 6 well<br /> *Lipofectamine(in uL) = DNA(in ug) * 2.5<br /> *OptiMEM(in uL, equal volume for DNA and Lipofectamine) = Lipofectamine(in uL) * 25<br /> <br /> ==Protocol (6 well dish)==<br /> #Plate cells and grow to ~70% confluence in 2 mL media without antibiotics (DMEM with 10% FBS, no PSG; use 1 mL for 12 well plate, and use half the amount of DNA)<br /> #For each well prepare one tube for DNA and one tube for Lipofectamine:<br /> ##OptiMEM plus required total amount of DNA.<br /> ##OptiMEM plus required total amount of Lipofectamine.<br /> ##Incubate ~5 minutes at room temperature.<br /> ##Combine the equal volumes of OptiMEM/DNA and OptiMEM/Lipofectamine.<br /> ##Incubate ~20 min at room temperature.<br /> #Add DNA/Lipofectamine complexes (~500 uL) to the 2 mL of media on the cells<br /> #Gently rock plate to mix<br /> #After ~4h aspirate media and re-feed cells with normal media. (It is usually possible to leave overnight). <br /> #Examine the cells the next day.<br /> <br /> Protocol adapted from [http://tools.invitrogen.com/content/sfs/manuals/lipofectamine2000_man.pdf Product Manual]</div>

Davebrid