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[DESCRIPTION]
<p>Procedure for DNA extractions using PowerWater DNA extraction kit from filtered LEO seepage water samples and prelimnary QC by nanodrop</p>

[BEFORE_START]
<p><strong>A) Identify samples to extract</strong></p>
<p>Determine <span style="text-decoration: underline;">number of samples to extract (n)</span></p>
<p>Open Google Drive spreadsheet: LEO seepage microbial communities - DNA extraction log</p>
<p>Itentify next n samples to be extracted - following extraction order</p>
<p>Make table or print from google drive with the following information</p>
<ul>
<li>Extraction number</li>
<li>Sample number</li>
<li>Sample ID</li>
<li>Full sample description (sample date/time, slope, replicate)</li>
</ul>
<p><strong>B) Gather and prepare supplies (inside of DNA extraction cabinet)</strong></p>
<p>Pipetters (P1000 and P200)</p>
<p>Pipette tips (filter tips)</p>
<p>Sharpie</p>
<p>Microtube rack</p>
<p>n x 1 Bead tubes </p>
<p>n x 5 Collection tubes</p>
<p>n x 1 Spin filter tubes</p>
<p> </p>
<p>Label tubes with Sample Number on top of the lid (NOT the same as extraction number)</p>
<p>Label FINAL collection tube on outside (1 for each sample), label with Sample Number (41), Sample ID (<span data-sheets-userformat="{&quot;2&quot;:4861,&quot;3&quot;:[null,0],&quot;5&quot;:{&quot;1&quot;:[{&quot;1&quot;:2,&quot;2&quot;:0,&quot;5&quot;:[null,2,0]},{&quot;1&quot;:0,&quot;2&quot;:0,&quot;3&quot;:3},{&quot;1&quot;:1,&quot;2&quot;:0,&quot;4&quot;:1}]},&quot;6&quot;:{&quot;1&quot;:[{&quot;1&quot;:2,&quot;2&quot;:0,&quot;5&quot;:[null,2,0]},{&quot;1&quot;:0,&quot;2&quot;:0,&quot;3&quot;:3},{&quot;1&quot;:1,&quot;2&quot;:0,&quot;4&quot;:1}]},&quot;7&quot;:{&quot;1&quot;:[{&quot;1&quot;:2,&quot;2&quot;:0,&quot;5&quot;:[null,2,0]},{&quot;1&quot;:0,&quot;2&quot;:0,&quot;3&quot;:3},{&quot;1&quot;:1,&quot;2&quot;:0,&quot;4&quot;:1}]},&quot;8&quot;:{&quot;1&quot;:[{&quot;1&quot;:2,&quot;2&quot;:0,&quot;5&quot;:[null,2,0]},{&quot;1&quot;:0,&quot;2&quot;:0,&quot;3&quot;:3},{&quot;1&quot;:1,&quot;2&quot;:0,&quot;4&quot;:1}]},&quot;9&quot;:0,&quot;10&quot;:2,&quot;12&quot;:0,&quot;15&quot;:&quot;arial,sans,sans-serif&quot;}" data-sheets-value="{&quot;1&quot;:2,&quot;2&quot;:&quot;Dec4-2030-W-2&quot;}">Dec4-2030-W-2</span>), your initials, and the date (you can do this with the label maker, 2 rows, 9 pt font)</p>
<p>Label bead tube with sample ID if not extracting right away. </p>
<p> </p>
<p><span style="text-decoration: underline;">Place all items in the DNA extraction cabinet, and sterilize by turning on uv lamp for 15 min</span> (twist dial completely, door must be closed)</p>
<p> </p>
<p><strong>C) Gather weighing supplies (room 202K)</strong></p>
<p>Rubber mallet</p>
<p>Ethanol</p>
<p>Kimwipes</p>
<p>Spatulas</p>
<p>Spare sample baggie </p>
<p> </p>
<p><strong>D) Prepare equipment (outside of DNA extraction cabinet)</strong><br />Timer<br />(+4°C) Tube rack in fridge<br />(+55°C) Heating block</p>
<p>Microcentrifuge </p>
<p>Centrifuge </p>
<p>Vortex next to hood</p>
<p>Vortex and adapter for bead tubes</p>
<p>Solutions PW1-PW6 from PowerWater DNA extraction kit </p>
<p>n x 1 50 mL conical tube (to pack bead tube in for centrifugation, do not have to be sterile or labeled)</p>
<p> </p>
<p><em>*Note: Three solutions are mislabeled by the manufacturer (write relevant PW# on top of lid)</em></p>
<p><em>PW2 = IRS (inhibitor removal solution)</em></p>
<p><em>PW5 = Ethanol</em></p>
<p><em>PW6 = EB (elution buffer)</em></p>
<p> </p>
<p><strong>Notes: </strong></p>
<p>Always minimize the amount of time a freezer is open by removing the box of interest, closing the freezer, adding/removing sample, and then reopening the freezer</p>
<p> </p>
<p>Samples must always be balanced in centrifuge rotor!</p>
<p> </p>
<p>Avoid passing your apendages over any open container by adjusting the position of your work items and your body. Store caps upside down on counters when remove - like this: U</p>
<p> </p>

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