https://eln.excellgen.com/index.php?action=history&feed=atom&title=Calcium_phosphate_transfection_protocol2 2026-10-18T06:10:46Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Calcium_phosphate_transfection_protocol2&diff=1133&oldid=prev 2011-06-22T22:43:01Z

<p>Created page with &quot;1. Detech cells with 0.05% tryp­sin–eth­yl­ene­di­amine­tet­ra­ace­tic acid (EDTA) treat­ment.&lt;br&gt; 2. Stop trys­i­ni­za­tion by DMEM–10% FCS.&lt;br&gt;3. Remove trys...&quot;</p> <p><b>New page</b></p><div>1. Detech cells with 0.05% tryp­sin–eth­yl­ene­di­amine­tet­ra­ace­tic acid (EDTA) treat­ment.&lt;br&gt;<br /> <br /> 2. Stop trys­i­ni­za­tion by DMEM–10% FCS.&lt;br&gt;3. Remove trysin by cen­tri­fu­ga­tion.&lt;br&gt;<br /> <br /> 4. Make Ca-P complex solu­tion using 2.2 M CaCl2 and 2X Hepes-buffered solu­tion [HBS] with ODN/siR­NA (200–250 nM).&lt;br&gt;<br /> <br /> 5. Add 70 ul of –cal­cium phos­phate pre­cip­i­tate to 2.5 X 10^5 cell sus­pension<br /> <br /> 6. Incubate for 25 min with gen­tle shak­ing every 5 min.<br /> <br /> 7. Add 1 ml of 10% FCS–DMEM,&amp;nbsp; plate cells in the serum-con­tain­ing medium for 3 to 4 h at 37 °C for adher­ence.<br /> <br /> 8. Remove medium, wash cells with PBS, add complete medium<br /> <br /> [[Category:Protocols]]&lt;br&gt;&lt;br&gt;</div>

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