<ol type="a">

<li> Lyse the resuspended cells by adding 200 µL of the Lysis Solution </li>

<li> Mix the contents by gentle inversion (6-8 times) until the mixture becomes clear and viscous (Don’t vortex) </li>

<li> Do not allow the lysis reaction to exceed 5 minutes </li>

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<li> Precipitate the cell debris by adding 350 µL of the Neutralization/Binding Solution </li>

<li> Gently invert the tube 4-6 times </li>

<li> Pellet the cell debris by centrifuging at ≥ 12,000 x g or maximum speed for 10 minutes </li>

<li> Cell debris, proteins, lipids, SDS, and chromosomal DNA should fall out as a cloudy, viscous precipitate. </li>

<li> If the supernatant contains a large amount of floating particulates after centrifugation, re-centrifuge the supernatant </li>

<ol type="a">

<li> Insert a GenElute Miniprep Binding Column into a provided micro-centrifuge tube, if not already assembled </li>

 

<li> Add 500 µL of the Column Preparation Solution to each miniprep column and centrifuge at ≥ 12,000 x g for 30 seconds to 1 minute </li>

<li> Discard the flow-through liquid </li>

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<li> Transfer the cleared lysate from step 3 to the column prepared in step 4 and centrifuge at ≥ 12,000 x g for 30 seconds to 1 minute. </li>

<li> Discard the flow-through liquid </li>

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<li> Add 750 µL of the diluted Wash Solution to the column </li>

<li> Centrifuge at ≥ 12,000 x g for 30 seconds to 1 minute </li>

<li> Discard the flow-through liquid and centrifuge again at maximum speed for 1-2 minutes without any additional Wash Solution to remove excess ethanol </li>

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<li> Transfer the column to a fresh collection tube. Add 100 µL of Elution Solution or molecular biology reagent water to the column </li>

<li> For DNA sequencing and other enzymatic application, use water or 5 Mm Tris-HCL, Ph 8.0, as an eluant </li>

<li> Centrifuge at ≥ 12,000 x g for 1 minute </li>

<li> Use immediately or store at -20˚C </li>