TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>The SpinSmart PCR purification and gel extraction technologies utilize a lysis buffer containing chaotropic salts that allow DNA to bind to a silica membrane. Binding buffer PCR 1 is added to a PCR reaction or agarose gel slice; the mixture is subsequently loaded directly onto SpinSmart PCR Columns.  Salts, enzymes, and other soluble components are washed away with ethanolic PCR 2 Wash buffer. Purified DNA is eluted using PCR 3 Elution buffer (5 mM Tris/HCl, pH 8.5). </p>

[BEFORE_START]
<p> </p>
<p>The following should be prepared before starting any SpinSmart PCR purification or gel extraction protocols:</p>
<p> </p>
<p>Add the indicated volume of 96-100% ethanol to PCR 2 Wash Buffer Concentrate.</p>
<p> </p>
<table>
<tbody>
<tr>
<td rowspan="2">
<p>PCR 2 Wash Buffer</p>
<p>(Concentrate)</p>
</td>
<td style="text-align: center;"><strong>50 preps</strong></td>
<td style="text-align: center;"><strong>250 preps</strong></td>
</tr>
<tr>
<td style="text-align: center;">2 x 6 ml<br />add 24 ml 96-100%<br />EtOH to each bottle</td>
<td style="text-align: center;">40 ml<br />add 160 ml 96-100%<br />EtOH</td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p> </p>
<p> </p>

[GUIDELINES]
<p><strong>Components </strong></p>
<p>SpinSmart PCR Purification and Gel Extraction Kit Contents</p>
<p><br /><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/g9qcz36.png" data-src="https://s3.amazonaws.com/pr-journal/g9qcz36.png" data-ofn="Screen Shot 2016-12-14 at 9.34.16 AM.png" /> </p>
<p> </p>
<p><strong>Equipment and reagents to be supplied by user</strong></p>
<p> </p>
<p><strong>Consumables</strong></p>
<p>96 - 100% ethanol</p>
<p>1.5 ml microcentrifuge tubes</p>
<p>Disposable pipette tips</p>
<p> </p>
<p><strong>Equipment</strong></p>
<p> </p>
<p>Manual pipettors</p>
<p>Centrifuge for microcentrifuge tubes</p>
<p>Heating block</p>
<p>Vortex mixer</p>
<p>Personal protection equipment (lab coat, gloves, goggles) </p>
<p> </p>
<p> </p>
<p><strong>The SpinSmart PCR Purification and Gel Purification Procedures </strong></p>
<p> </p>
<p>The SpinSmart PCR purification and gel extraction technologies utilize a lysis buffer containing chaotropic salts that allow DNA to bind to a silica membrane. Binding buffer PCR 1 is added to a PCR reaction or agarose gel slice; the mixture is subsequently loaded directly onto <strong>SpinSmart PCR Columns</strong>.  Salts, enzymes, and other soluble components are washed away with ethanolic PCR 2 Wash buffer. Purified DNA is eluted using PCR 3 Elution buffer (5 mM Tris/HCl, pH 8.5). </p>
<p> </p>
<p><strong>SpinSmart PCR Kit Specifications </strong></p>
<p> </p>
<p>SpinSmart PCR kits are designed for DNA purification from TAE/TBE agarose gels and for the direct purification of PCR* products. </p>
<p> </p>
<p>SpinSmart PCR buffers are formulated to completely remove primers from PCR* reactions. Small double-stranded DNA fragments still remain bound and are purified with high efficiency. </p>
<p> </p>
<p>SpinSmart PCR kits will effectively purify DNA fragments from detergent-rich PCR* reaction buffers.</p>
<p> </p>
<p>DNA absorption to the membrane is pH-dependent. TAE standard gels or reaction mixtures with pH 6-8 should be used for best results. </p>
<p> </p>
<p>Both standard and low melting agarose gels can be used. </p>
<p> </p>
<p>SpinSmart PCR purified DNA fragments are ready to use in downstream applications like automated fluorescent DNA sequencing, PCR (PCR is patented by Roche Diagnostics), ligation reactions, or other types of enzymatic manipulation. </p>
<p> </p>
<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/g9rcz36.png" data-src="https://s3.amazonaws.com/pr-journal/g9rcz36.png" data-ofn="Screen Shot 2016-12-14 at 9.35.28 AM.png" /></p>
<p> </p>
<p><strong>Removal of small DNA fragments and primer-dimers </strong></p>
<p> </p>
<p><strong>Spin Smart PCR i</strong>s specially formulated to remove unused, single stranded primers while effectively purifying PCR products down to 60 bp. In some cases, a PCR reaction may yield unwanted small double stranded fragments, such as primer-dimers or small PCR products resulting from unspecific annealing. SpinSmart PCR offers a simple method to remove these products that can interfere with your downstream sequencing or cloning applications. </p>
<p> </p>
<p>By simply diluting PCR 1 Binding Buffer with sterile water, you can decrease the ability of small DNA fragments to bind to the membrane without compromising larger fragment recovery.  A simple dilution series should be tested, ranging from 1:1 – 1:9 (PCR 1:H2O) in order to determine the appropriate cutoff range for your reaction.  As you approach the 1:9 dilution, the larger fragment recovery will sequentially decrease as well. </p>
<p> </p>
<p><em>Rule of Thumb</em>: The smaller the fragment you wish to exclude, the less you will need to dilute the PCR 1 Binding Buffer.</p>
<p> </p>
<p> </p>
<p><strong>Elution procedures</strong></p>
<p> </p>
<p>DNA should be eluted using the PCR 3 Elution Buffer.  If necessary, sterile water or other low salt elution buffers may be used, however the pH must be in the range of 7.0 – 8.5 for optimal recovery.   </p>
<p> </p>
<p>Typical recovery of 70-95% can be obtained with DNA fragments between 50bp -10kbp with an elution volume of 15 µl. For larger amounts of DNA (5-15 µg of DNA; from PCR* reactions &gt; 100 µl or gel slices &gt; 200 mg), elution with at least 50 µl of PCR 3 Elution Buffer is recommended.</p>
<p> </p>
<p>Pre-warmed PCR 3 Elution Buffer can improve the yields of larger fragments (&gt; 5-10 kbp). Add pre-warmed PCR 3 Elution Buffer (70°C) to the membrane, and incubate for 1-2 minutes, then centrifuge as directed in the standard protocol.</p>
<p><br /><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/g9scz36.png" data-src="https://s3.amazonaws.com/pr-journal/g9scz36.png" data-ofn="Screen Shot 2016-12-14 at 9.37.00 AM.png" /></p>
<p> </p>
<p><strong>Storage and preparation of solutions</strong></p>
<p> </p>
<p> </p>
<p>PCR 1 Wash Buffer contains chaotropic salt. Wear gloves and goggles!</p>
<p> </p>
<p>SpinSmart PCR kit components should be stored at room temperature and are stable for up to one year.</p>
<p> </p>
<p>The following should be prepared before starting any SpinSmart PCR purification or gel extraction protocols:</p>
<p> </p>
<p>Add the indicated volume of 96-100% ethanol to PCR 2 Wash Buffer Concentrate.</p>
<p> </p>
<table>
<tbody>
<tr>
<td rowspan="2">
<p>PCR 2 Wash Buffer</p>
<p>(Concentrate)</p>
</td>
<td style="text-align: center;"><strong>50 preps</strong></td>
<td style="text-align: center;"><strong>250 preps</strong></td>
</tr>
<tr>
<td style="text-align: center;">2 x 6 ml<br />add 24 ml 96-100%<br />EtOH to each bottle</td>
<td style="text-align: center;">40 ml<br />add 160 ml 96-100%<br />EtOH</td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>Safety instructions – risk and safety phrases</strong></p>
<p>The following components of the SpinSmart PCR kits contain hazardous materials. <br /><em>Wear gloves and goggles and follow the safety instructions given in this section!</em></p>
<p> </p>
<p><strong><em><img class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/g9ucz36.png" data-src="https://s3.amazonaws.com/pr-journal/g9ucz36.png" data-ofn="Screen Shot 2016-12-14 at 9.41.49 AM.png" /></em></strong></p>
<p><strong><em>Risk Phrases</em></strong> </p>
<p><em>R 20/21/22 Harmful by inhalation, in contact with the skin and if swallowed</em><strong><em> <br />Safety Phrases</em></strong></p>
<p><em>S 13 Keep away from food, drink and animal feedstuffs</em></p>
<p> </p>
<p><em>∗ Label not necessary, if quantity below 125 g or ml (according to 67/548/EEC Art. 25, 1999/45/EC Art. 12 and German GefStoffV § 42 and TRGS 200 7.1) <br /></em></p>
<p> </p>
<p><strong>Troubleshooting</strong></p>
<p>For troubleshooting see the product manual:</p>
<p><a href="https://www.denvillescientific.com/Files/Files/Denville/ProductDocs/CM-510-50_and_CM-510-250_manual.pdf" target="_blank">https://www.denvillescientific.com/Files/Files/Denville/ProductDocs/CM-510-50_and_CM-510-250_manual.pdf</a></p>
<p> </p>
<p> </p>

[STEPS]
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