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[DESCRIPTION]
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<div class="column">The <strong>DNA Clean &amp; Concentrator™-5 (DCC™-5)</strong> provides a hassle-free method for the rapid purification and concentration of high-quality DNA from PCR, endonuclease digestions, cell lysates, and other impure DNA preparations. It can also be used for post-RT cDNA clean-up and purification of sequencing-ready DNA from M13 phage. </div>
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[BEFORE_START]
<p><strong>Buffer Preparation</strong></p>
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<p><em>Before starting:</em> Add 24 ml 100% ethanol (26 ml 95% ethanol) to the 6 ml<strong> DNA Wash Buffer</strong> concentrate. Add 96 ml 100% ethanol (104 ml 95% ethanol) to the 24 ml <strong>DNA Wash Buffer</strong> concentrate.</p>
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[GUIDELINES]
<p><strong>Product Contents</strong></p>
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<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gthc2w6.png" width="450" data-src="https://s3.amazonaws.com/pr-journal/gtgc2w6.png" data-ofn="Screen Shot 2016-11-25 at 10.38.44 AM.png" /></p>
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<p>Integrity of kit components is guaranteed for up to one year from date of purchase. Reagents are routinely tested on a lot-to-lot basis to ensure they provide the highest performance and reliability.</p>
<p><sup>1</sup> Ethanol must be added prior to use as indicated on the <strong>DNA Wash Buffer</strong> label.</p>
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<p><strong>Specifications</strong></p>
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<p><strong>DNA Purity</strong> – High-quality DNA (A<sub>260</sub>/A<sub>280</sub> &gt;1.8) ideal for ligation, sequencing, labeling, PCR, microarray, transfection, transformation, and restriction digestion procedures.</p>
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<p><strong>DNA Size Limits</strong> – From ~50 bp to 23 kb.</p>
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<p><strong>DNA Recovery</strong> – Typically, up to 5 μg total DNA per column can be eluted into as little as 6 μl of low salt DNA Elution Buffer or water. For DNA 50 bp to 10 kb, the recovery is 70-90%. For DNA 11 kb to 23 kb, the recovery is 50-70%.</p>
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<p><strong>Sample Sources</strong> – DNA from enzymatic reactions (e.g., PCR, restriction endonuclease digestions), plasmid preparations, and impure preparations.</p>
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<p><strong>Product Detergent Tolerance</strong> – ≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤ 0.1% SDS.</p>
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<p><strong>Product Description</strong></p>
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<p>The <strong>DNA Clean &amp; Concentrator™-5 (DCC™-5)</strong> provides a hassle-free method for the rapid purification and concentration of high-quality DNA from PCR, endonuclease digestions, cell lysates, and other impure DNA preparations. It can also be used for post-RT cDNA clean-up and purification of sequencing-ready DNA from M13 phage. Simply add the specially formulated <strong>DNA Binding Buffer</strong> to your sample and transfer the mixture to the supplied <strong>Zymo-Spin™ Column</strong>. There is no need for organic denaturants or chloroform. Instead, the product features Fast-Spin column technology to yield DNA that is free of salts and contaminants in just 2 minutes. The purified DNA is ideal for DNA ligation, sequencing, labeling, PCR, microarray, transfection, transformation, and restriction digestion procedures.</p>
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<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gtic2w6.png" width="450" data-src="https://s3.amazonaws.com/pr-journal/gtic2w6.png" data-ofn="Screen Shot 2016-11-25 at 10.43.10 AM.png" /></p>
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<p><strong>Available Formats</strong></p>
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<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gtkc2w6.png" width="450" data-src="https://s3.amazonaws.com/pr-journal/gtjc2w6.png" data-ofn="Screen Shot 2016-11-25 at 10.44.11 AM.png" /></p>
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<p><strong>Typical DCC™ Applications</strong></p>
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<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gtnc2w6.png" width="450" data-src="https://s3.amazonaws.com/pr-journal/gtmc2w6.png" data-ofn="Screen Shot 2016-11-25 at 10.45.21 AM.png" /></p>
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<li>For purification of short DNA or RNA oligonucleotides ≥ 16 nt, use the <strong>Oligo Clean &amp; Concentrator (D4060, D4061)</strong>.</li>
<li> For ChIP (Chromatin Immunoprecipitation) sample cleanup, use the <strong>ChIP DNA Clean &amp; Concentrator (D5201,</strong><strong>D5205)</strong> for high quality DNA from any step in a standard ChIP protocol.</li>
<li> For post-cycle sequencing samples, use the <strong>ZR Sequencing DNA Clean-up Kit (D4050, D4051)</strong> for dye blob elimination.</li>
<li> For samples containing PCR inhibitors, use the <strong>OneStep™ PCR Inhibitor Removal Kit (D6030, D6035).</strong></li>
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<p><strong>Buffer Preparation</strong></p>
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<p><em>Before starting:</em> Add 24 ml 100% ethanol (26 ml 95% ethanol) to the 6 ml<strong> DNA Wash Buffer</strong> concentrate. Add 96 ml 100% ethanol (104 ml 95% ethanol) to the 24 ml <strong>DNA Wash Buffer</strong> concentrate.</p>
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<p><strong>Troubleshooting</strong></p>
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<p><strong>Low Recovery</strong></p>
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<li><strong>Improperly Prepared/Stored DNA Wash Buffer</strong></li>
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<p>Make sure ethanol has been added to the DNA Wash Buffer concentrate. Cap the bottle tightly to prevent evaporation over time.</p>
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<p><strong>Addition of DNA Elution Buffer</strong><br />Add elution buffer directly to the column matrix, not to the walls of the column. Elution buffer requires contact with the matrix for at least 1 minute for large DNA ≥ 10kb.</p>
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<p><strong>Incomplete Elution</strong></p>
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<p>DNA elution is dependent on pH, temperature, and time. For large genomic DNA (≥ 50 kb), apply heated elution buffer (60-70 °C) to the column and incubate for several minutes prior to elution.</p>
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<p>Sequential elutions may be performed for quantitatively higher recovery but lower final DNA concentration. This is recommended for DNA ≥ 10 kb.</p>
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<div class="column"><strong>Low A260/A230 ratio</strong><br />
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<li><strong>Column tip contaminated</strong></li>
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<p>When removing the column from the collection tube, be careful that the tip of the column does not come into contact with the flowthrough. Trace amounts of salt from the flowthrough can contaminate a sample resulting in a low A<sub>260</sub>/A<sub>230</sub> ratio. Ethanol contamination from the flowthrough can also interfere with DNA elution. Zymo-Spin™ columns are designed for complete elution with no buffer retention or carryover (see below).</p>
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<p><img class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gtdc2w6.png" width="450" data-src="https://s3.amazonaws.com/pr-journal/gtdc2w6.png" data-ofn="Screen Shot 2016-11-25 at 10.25.39 AM.png" /></p>
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<p><strong>Following Clean-up with the DCCTM, Multiple Bands Appear in an Agarose Gel</strong></p>
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<li><strong>Acidification of DNA Loading Dye</strong></li>
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<p>Most loading dyes do not contain EDTA and will acidify (pH ≥ 4) over time due to some microbial growth. This low pH is enough to cause DNA degradation. Therefore, if water is used to elute the DNA, 6X Loading Dye containing 1 mM EDTA is recommended.</p>
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<p>--------------------------- </p>
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<p><strong>Selected Citations</strong></p>
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<p>Li, N. (2010). Whole genome DNA methylation analysis based on high throughput sequencing technology. Methods, 52 (3), 221-232.</p>
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<p>Lee, EJ. (2011). Targeted bisulfite sequencing by solution selection and massively parallel sequencing. Nucleic Acids Research, 39(19),</p>
<p>e127, doi:10.1093/nar/gkr598</p>
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<p>Papageorgiou, EA. (2009). Sites of differential DNA methylation between placenta and peripheral blood. Am J Pathol, 174 (5), 1609-1618.</p>
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<p>Ferguson, A.A. et al. (2009). Retrofitting ampicillin resistant vectors by recombination for use in generating C. elegans transgenic animals by bombardment. Plasmid, 62, 140-145.</p>
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