TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol is a modified version of the MoBio PowerPlant Pro 96 well DNA extraction kit for plant and lichen tissues that have been stored in CTAB buffer.  Prior to being placed in CTAB, photosynthetic tissues from plants and lichens were surface-sterilized with sequential washes in 95% EtOH, 0.5% sodium hypochlorite, and 70% EtOH and air dried under sterile conditions.  Twenty-four tissue pieces per host were placed in 750 uL of CTAB buffer.  Four tubes per host individual were sampled for a total of 96 tissue pieces. </p>

[BEFORE_START]
<p><strong>You will need the following reagents and equipment for the protocol. </strong></p>
<p> </p>
<p><strong>Reagents<br /></strong></p>
<p><em>-DNase Away</em></p>
<p><em>-Stainless Steel Beads, 3.2 mm (Next Advance Cat # SSB32)</em></p>
<p><em>-Stainless Steel Bead blend, 0.9-2.0mm (NextAdvance Cat # SSB14B)</em></p>
<p><em>-Zirconium Oxide Beads, 2.0 mm (Next Advance Cat # ZRoB20)</em></p>
<p><em>-Liquid Nitrogen</em></p>
<p><em>-Molecular grade 100% EtOH</em></p>
<p><em>-MoBio PowerPlant Pro 96 well DNA extraction kit (each kit processes 4 96 well plates)</em></p>
<p><strong> </strong></p>
<p><strong>Equipment:</strong></p>
<p><em>-Laminar flow hood</em></p>
<p><em>-Lyophilizer</em></p>
<p><em>-Centrifuge capable of handling 24 1.5-2.0 mL tubes</em></p>
<p><em>-Centrifuge capable of spinning 96 well (deep) plates from MoBio at 4,000 x g.</em></p>
<p><em>-Next Advance Bullet Blender Storm or comparable bead beater</em></p>
<p><em>-Heat block and water bath at 65C</em></p>
<p><em>-Parafilm</em></p>
<p><em>-Sterile needle</em></p>
<p><em>-50 mL Falcon tubes (DNA/RNA free)</em></p>
<p><em>-1000 uL filter tips</em></p>
<p><em>-200 uL filter tips</em></p>
<p><em>-300 uL filter tips (best for transferring supernatant with multichannel pipette)</em></p>
<p><em>-Aluminum sealing tape</em></p>
<p><em>-Reagent reservoirs</em></p>
<p><em>-Two multichannel pipettes.  One capable of small volumes (&lt;100 uL) and another for pipetting large volumes (&gt;200 uL)</em></p>
<p><em>-Two freezer blocks (-20C)</em></p>

[GUIDELINES]
<ul>
<li>Gloves must be worn at all times. Change gloves frequently and decontaminate often with DNA Away.</li>
<li>Pipetting must be done extremely carefully to minimize the risk of aerosols that can easily cause contamination between samples.</li>
<li>Use only unopened, sterile, aerosol-resistant pipette tips (filter tips) to minimize contamination of the pipette shaft and your samples. Clean the body of the pipettes regularly with DNA Away.</li>
<li>Discard pipette tips after each use to avoid cross-contamination. Eject pipette tips carefully to prevent aerosol formation or other splashing.</li>
<li>Remember to always clean the hood and any equipment with DNA Away and decontaminate where possible with UV for 30 minutes prior to use to avoid cross-contamination.</li>
<li>The pipettes used need to be calibrated annually.</li>
<li>Be careful not to touch the inside lid of tubes as it will cross-contaminate samples.</li>
</ul>

[STEPS]
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