https://eln.excellgen.com/index.php?action=history&feed=atom&title=Plasmid_Purification_by_Aqueous_Two_Phase_SystemPlasmid Purification by Aqueous Two Phase System - Revision history2026-10-18T04:03:18ZRevision history for this page on the wikiMediaWiki 1.44.0https://eln.excellgen.com/index.php?title=Plasmid_Purification_by_Aqueous_Two_Phase_System&diff=15156&oldid=prevPaul.xu: Created page with "Spin down cells.<br/>Add P1, votex to resuspend bacteria completely<br/>Add P2 to lyse cells. swirl to mix, do not votex<br/>Add P3 to neutralize<br/>Add 0.7 volume of isoprop..."2016-08-01T16:34:37Z<p>Created page with "Spin down cells.<br/>Add P1, votex to resuspend bacteria completely<br/>Add P2 to lyse cells. swirl to mix, do not votex<br/>Add P3 to neutralize<br/>Add 0.7 volume of isoprop..."</p>
<p><b>New page</b></p><div>Spin down cells.<br/>Add P1, votex to resuspend bacteria completely<br/>Add P2 to lyse cells. swirl to mix, do not votex<br/>Add P3 to neutralize<br/>Add 0.7 volume of isopropanol, incubate on ice for 90 min<br/>Spin for 20 min at 4ºC.<br/>Dissolve pellet in 3.5 ml of 10 mM Tris - HCl buffer (pH 8.0), or TE.<br/>Add solid ammonium sulphate to the solution to a concentration of 2.5 M.<br/>Incubate on ice 15 min.<br/>Spin for 20 min &nbsp;at 4ºC to remove RNA.&nbsp;<br />
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Purification using PEG-(NH4)2SO4 two phase system<br/>20% w/w in a system composed of 34% w/w PEG 600 and 7% w/w (NH4)2SO4,&nbsp;<br/>or &nbsp;PEG 600 (19.0% w/w) with 12.4% sodium citrate and 4.1% (NH4)2SO4<br />
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Purification using HIC<br/>Load supernatant to a a Phenyl Sepharose 6 Fast Flow column (XK 16/20 column) with a 14 cm bed height pre-equilibrated with 1.5 M<br/>ammonium sulphate in 10 mM Tris-HCl (pH 8.0), &nbsp;flow rate is 1 ml/min.&nbsp;<br/>Collect flow through (contains plasmid DNA)<br/>Elute genomic DNA and RNA off column using water.<br />
[[Category:QC]]</div>Paul.xu