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[BEFORE_START]
<p>Please wear at least the minimum required personal protective equipment.</p>
<p>Ensure that all necessary kit components are available as well as user-supplied consumables.</p>
<p>Clean all working surfaces, pipettes, and pens to remove DNA contamination.</p>

[GUIDELINES]
<p><strong><a href="https://mobio.com/media/wysiwyg/pdfs/protocols/27000-4-KF.pdf" target="_blank" rel="noopener noreferrer">QIAGEN® MagAttract® PowerSoil® DNA KF Kit (384) (Optimized for KingFisher)</a></strong></p>
<p> </p>
<p>The Knight lab has transitioned to the QIAGEN® MagAttract® PowerSoil® DNA KF Kit (384) (Optimized for KingFisher). We have validated a variety of sample types to ensure reproducibility when compared to MoBio PowerSoil Extraction Kit. This transition occurred to increase efficiency and reduce DNA extraction time from 6-8 hours to 3-4 hours.</p>
<p> </p>
<p>The protocol is followed as QIAGEN® recommends, with an added 10-minute water bath incubation at 65°C after the Lysis buffer addition, and retention of the 10-minute incubation at 4°C after lysate addition to IR Solution.</p>
<p> </p>
<p>The new kit can be implemented on the epMotion using a magnetic bead plate adapter for the epMotion. However, it does not reduce the amount of time the extraction takes by a significant amount: tests showed that the new kit on the epMotion took longer than the old kit. Comparison showed both the old and new kit performed well on the epMotion.</p>
<p> </p>
<p><a href="https://mobio.com/products/dna-isolation/soil/powersoilr-htp-96-well-soil-dna-isolation-kit.html" target="_blank" rel="noopener noreferrer"><strong>QIAGEN® MagAttract® PowerSoil® DNA KF Kit (384)</strong></a></p>
<p> </p>
<p>The QIAGEN® DNeasy® PowerSoil® Kit (formerly: MoBio PowerSoil DNA Isolation Kit; QIAGEN® catologue: 12888-100) is still used if KingFisher instrumentation is not available, or if the sample set contains less than 48 samples.</p>
<p> </p>
<p><strong>Items included in the extraction kit (for one 96 well extraction)</strong></p>
<ul>
<li>(1) Bead plate</li>
<li>(2) 1.0ml collection plates</li>
<li>Sealing Tape</li>
<li>Labeled solutions</li>
</ul>
<p> </p>
<p><strong>Important Considerations </strong></p>
<p> </p>
<ol>
<li>Normal diameter 1 ml pipet tips are too large for some of the pipetting steps. To get around this problem we use a Rainin™ 1000 ul 8-channel pipette with extended length filtered tips (Rainin™ RT-LTS-A-1000µL-/F/X-768/8, catologue: <span data-sheets-value="{&quot;1&quot;:2,&quot;2&quot;:&quot;30389223&quot;}" data-sheets-userformat="{&quot;2&quot;:513,&quot;3&quot;:[null,1],&quot;12&quot;:0}">30389223</span>).</li>
<li>Make sure that all of the necessary consumables and reagents are in place before you start the extraction. Remember, each pipetting step will require 1 box of 96 tips per plate.</li>
<li>We use presterilized reservoirs for pipetting steps carried out by epMotion® liquid handlers.</li>
<li>When setting up the mechanical lysis, it is important that the plate not rub against any surfaces of/in the plate shaker.</li>
<li>Make sure that the alpha-numeric grid is in the same orientation across all the plates.</li>
</ol>
<p><br /><strong>Next Steps</strong></p>
<ul>
<li><a href="https://www.protocols.io/view/emp-16s-rrna-amplification-protocol-nuudeww" target="_blank" rel="noopener noreferrer">16S rRNA Amplification Protocol</a></li>
<li><a href="https://www.protocols.io/view/emp-18s-rrna-amplification-protocol-nuvdew6?step=6" target="_blank" rel="noopener noreferrer">18S rRNA Amplification Protocol</a></li>
</ul>

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