https://eln.excellgen.com/index.php?action=history&feed=atom&title=Plasmid_MaxiPrep_Simple 2026-10-19T02:30:42Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Plasmid_MaxiPrep_Simple&diff=6523&oldid=prev 2012-10-13T03:40:49Z

<p></p> <p><b>New page</b></p><div>== Miniprep ==<br /> <br /> 1. Spin down cells 1.5-2 ml of bacterial cultures were pelleted at 6000-7000 rpm for 1 min.&lt;br&gt; 2. Remove culture medium.&lt;br&gt; <br /> <br /> 3. Add suspension buffer:&lt;br&gt; <br /> <br /> 5% sucrose, 20–50 mM EDTA, 50 mM Tris pH 8, 0.75 M NH4Cl, 0.5% IGEPAL CA-630 (or Triton X-100), lysozyme 100 µg/ml, and RNase A 25 µg/ml, 20–50 mM CaCl2.&lt;br&gt; <br /> <br /> 4.&amp;nbsp;Incubated at 65°C for 5 min.&lt;br&gt; 5. Transfer supernatant to another tube.&lt;br&gt; <br /> <br /> 6. Add 100-120 µl isopropanol,&amp;nbsp; mix and and spin at 7000 rpm for 10 min at RT.&lt;br&gt; <br /> <br /> 7. Discard supernatant. &lt;br&gt; <br /> <br /> 8. Wash with 70% ethanol.&lt;br&gt; <br /> <br /> 9. Remove ethanol.&lt;br&gt; <br /> <br /> 10. Dissolve in 50 ul TE.&lt;br&gt; <br /> <br /> &lt;br&gt;&amp;nbsp; 100X enzyme stock containing 10 mg/ml lysozyme and 2.5 mg/ml of RNase A prepared in 50% glycerol and 50 mM Tris pH 8 was stored at −20°C and used repeatedly.&lt;br&gt; <br /> <br /> <br /> <br /> == Maxiprep ==<br /> <br /> <br /> <br /> Ref:&lt;br&gt; <br /> <br /> http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023457?imageURI=info:doi/10.1371/journal.pone.0023457.t001&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Protocols]]</div>

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