One step 'miniprep' method for the isolation of plasmid DNA protocol

Solutions/reagents: overnight E.coli culture phenol : chloroform : isoamyl alcohol(25:24:1)  (phenol saturated with TE(10mM Tris, 7.5, 1mM EDTA) prior to mixing with chloroform and isoamylalcohol) isopropanol70% ethanol100 µl/ml RNAse Equipment: CentrifugeSterile 1.5-ml microcentrifuge tubes</li></ul> Steps: <ol> Measure out <font color=#357EC7>0.5 ml  of <font color=#357EC7>overnight E.coli culture into sterile 1.5-ml microcentrifuge tube (1). <font color = "#800517">We routinely grow our cells in 'standard 1' bacteriological media supplied by Merck, Germany. </li> Measure out <font color=#357EC7>0.5 ml  of <a href="#phenol : chloroform : isoamyl alcohol(25:24:1)" ><font color=#357EC7>phenol : chloroform : isoamyl alcohol(25:24:1) </a> into sterile 1.5-ml microcentrifuge tube (1). </li> Vortex the mixture for <font color=#357EC7>1 min . <font color = "#800517">Vortex at maximum speed. <font color = "#800517">Alternatively, vortex for 10 seconds and then transfer to eppendorf mixer or an over-the-top rotator for 5 minutes. </li> Centrifuge at a speed of <font color=#357EC7>12000 Xg for <font color=#357EC7>5 mins  at <font color=#357EC7><b><font color=#357EC7>room temperature  </b>. </li> Meanwhile: Set aside a fresh sterile 1.5-ml microcentrifuge tube (2). Call it Tube I. Measure out <font color=#357EC7>0.5 ml  of <font color=#357EC7>isopropanol into Tube I. </li> Add <font color=#357EC7>0.45 ml  of <font color=#357EC7>top aqueous phase obtained after centrifugation. Vortex the mixture for a few secs. Centrifuge at a speed of <font color=#357EC7>12000 Xg for <font color=#357EC7>5 mins  at <font color=#357EC7>room temperature , gently aspirate out the supernatant and discard it. <font color = "#800517">Addition of salt and cooling is unnecessary. </li> Add <font color=#357EC7>0.5 ml  of <font color=#357EC7>70% ethanol. <font color = "#800517">Add ethanol carefully to the side of the tube. Discard solution. </li> Repeat Step 7. Add <font color=#357EC7>100 µl/ml RNAse to solution. Resuspend pellet by vortexing/by shaking vigorously. <font color = "#800517">About 5-10ul of this DNA can now be cleaved with appropriate restriction enzyme(s) for analysis. </li> </ol> TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :<font color=#357EC7>~ 11 mins