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[DESCRIPTION]
<p>1µl Assay for detecting and measuring DNA, RNA &amp; Oligos.</p>

[GUIDELINES]
<p><strong>INTRODUCTION </strong></p>
<p>The patented NUCLEIC dotMETRIC™ kit provides a quick and accurate method for detecting and measuring DNA, RNA and oligonucleotides.  This system is an alternative to spectrophotometry; no expensive equipment or cuvettes are necessary.  NUCLEIC dotMETRIC™ uses as little as a microliter of your sample and provides permanent results in minutes.   </p>
<p> </p>
<p><strong><em>Two methods in one:</em></strong> NUCLEIC dotMETRIC™ offers two separate protocols for measuring nucleic acid concentration; the concentration of nucleic acid can be measured either by measuring the diameter of nucleic acid spots or comparing the color density of the nucleic acid spots with that of a set of known standards.</p>
<p> </p>
<p>To use NUCLEIC dotMETRIC™, dilute samples with dilution buffer, spot 1‐5 µl of each sample onto the NUCLEIC Test Strip, develop the strip with NUCLEIC dyes, wash and dry. Using the NUCLEIC dotMETRIC™ scale, dot diameter is measured for accurate determination of nucleic acid concentration. Alternatively, compare the color density of the nucleic acid spots with that of a set of known standard nucleic acid spots.  </p>
<p> </p>
<p>Samples in the range of 10µg/µl to 1ng/µl can be measured accurately regardless of the isolation method or storage buffer. Oligonucleotides as short as 16 bases, have successfully been measured with NUCLEIC dotMETRIC™ . Measurements are not affected by proteins. The lower detection limit of the system is 1‐2ng/µl making NUCLEIC dotMETRIC™ one of the most sensitive methods available for measuring nucleic acids.</p>
<p> </p>
<p><strong>ITEM(S) SUPPLIED</strong></p>
<table>
<tbody>
<tr style="height: 20px;">
<td style="height: 20px;"><strong>Description</strong></td>
<td style="height: 20px;"><strong>Cat. # 786‐60</strong></td>
<td style="height: 20px;"><strong>Cat. # 786‐61</strong></td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">NUCLEIC Test Strips</td>
<td style="height: 20px;">50</td>
<td style="height: 20px;">50</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">NUCLEIC Dye</td>
<td style="height: 20px;">30ml</td>
<td style="height: 20px;">30ml</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">Nucleic Acid Dilution Buffer</td>
<td style="height: 20px;">10ml</td>
<td style="height: 20px;">10ml</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">dotMETRIC™ Scale</td>
<td style="height: 20px;">1</td>
<td style="height: 20px;">1</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">dotMETRIC™ Standard</td>
<td style="height: 20px;">1</td>
<td style="height: 20px;">1</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">Application Board</td>
<td style="height: 20px;">1</td>
<td style="height: 20px;">1</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">Forceps</td>
<td style="height: 20px;">1</td>
<td style="height: 20px;">1</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">dotMETRIC™ Application Device</td>
<td style="height: 20px;">-</td>
<td style="height: 20px;">1</td>
</tr>
<tr style="height: 20px;">
<td style="height: 20px;">1µl Application Capillary</td>
<td style="height: 20px;">-</td>
<td style="height: 20px;">100 </td>
</tr>
</tbody>
</table>
<p> </p>
<p><strong>STORAGE CONDITIONS</strong></p>
<p>The kit is shipped at ambient temp.  Upon arrival, store it at room temperature.</p>
<p> </p>
<p><strong>ADDITIONAL ACCESSORIES</strong></p>
<p>The following items are also available separately:</p>
<p>• Application (Pipette) Tips [Cat #786‐64]</p>
<p>• dotMETRIC™ Spot Application Device [Cat #786‐63 ]</p>
<p>• 1µl Application Capillary [Cat #786‐23]</p>
<p>• Developing Trays [Cat #786‐24] </p>
<p> </p>
<p><strong>QUICK PROTOCOL</strong></p>
<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/d7dc3kn.png" data-src="https://s3.amazonaws.com/pr-journal/d7dc3kn.png" data-ofn="Screen Shot 2016-07-06 at 2.13.05 PM.png" /></p>
<p>1. Dilute sample in Dilution Buffer 1:1 to 1:10 fold.</p>
<p>2. Apply 1‐5µl sample on the Test Strip, either as a free drop or as point of contact capillary action, or both.</p>
<p>3. Develop the Test Strip and determine concentration.</p>
<p> </p>
<p><strong>SPOT APPLICATION DEVICE</strong></p>
<p>Due to the above‐indicated limitations, we strongly recommend using G‐Biosciences’ Spot Application Device. The Spot Application Device is easy to work with and generally makes application easy and gives better results</p>
<p> </p>
<p><em>Using Application Device for Applying Nucleic Acid Samples.  </em></p>
<p> </p>
<p><strong>1.</strong> Perform nucleic acid application in a well‐lit area and make sure that you can see<br />the solution rise and flow through the application capillaries.  <br /><strong>2.</strong> Use forceps for handling the application capillary tips. Place the application tip into<br />the solution mixed with the Dilution Buffer; the solution will rise up into the<br />capillary application tip and fill within 3‐5 seconds.<br /><strong>3.</strong> Position the application tips in the holes provided in the device and lower the tip<br />until the tip touches the membrane surface. As soon as the tip touches the<br />membrane, the solution will begin to flow in to the membrane.  <br /><strong>4.</strong> After the samples have been diffused into the membrane and the application tip is<br />empty of the solution, remove the application tip from the device. Same capillary<br />tip can be used for several applications.<br /><strong>5.</strong> For applying more than 1µl sample on each spot (2‐4µl), apply the sample multiple<br />times<br />NOTE:  Allowing the sample to dry before applying a second time is not necessary.</p>
<p> </p>
<p><strong>TROUBLESHOOTING </strong></p>
<p><strong>Problem</strong>: Sample concentrations below 0.03µg/µl. When nucleic acid concentrations fall below 0.03 µg/µl. The diameter of nucleic acid spots is no longer large enough to be measured with the dotMETRIC™ scale.</p>
<p><strong>Solution</strong>: The lower detection limit of nucleic acid in dotMETRIC™ application averages around 1ng/ µl. Take 1‐2 µl of sample and prepare serial dilutions in dilution buffer, such as x2, x4, x8, x16, x32, x64.fold. Use dotMetric™ application and spot 1 µl from each dilution. Develop the strip. For concentration determination find the last visible spot at the highest dilution. Multiply the dilution fold of the last visible spot by (1+0.1) ng/ul.</p>
<p> </p>
<p><strong>Problem</strong>: DNA solution does not flow easily into the Test Strip.</p>
<p><strong>Solution</strong>: This is due to large strings of DNA in the solution. Dilute the DNA solution further with the Dilution buffer to lower the DNA concentration below &lt;1µg / µl and after mixing, shear the DNA by vigorously vortexing, sonication, or pipetting.</p>
<p> </p>
<p><strong>Problem</strong>:  Dots are not a uniform circle.</p>
<p><strong>Solution</strong>:  Take care to maintain the pipette tip in a straight up position while the sample is drawn by capillary action of the NUCLEIC Strip. Samples generally take 5‐15 seconds to be absorbed depending on the concentration of nucleic acid. Alternatively, use Spot Application Device.  </p>
<p> </p>
<p><strong>Problem</strong>:  The dots are faint and/or grainy in appearance.  In some instances dots are surrounded by a fuzzy  'halo'.</p>
<p><strong>Solution</strong>:  These problems are usually caused by high concentrations of impurities in your samples (i.e., salts, phenol, organic solvents). Samples with high concentrations of nucleic acids can be diluted further with the appropriate dilution buffer and measured again.    A second solution is to purify your samples using GET™ CLEAN DNA (Cat. # 786‐356 or 786‐357) or ethanol precipitation.  Removing impurities from your samples is also likely to improve your success with any down‐stream applications.</p>
<p> </p>
<p><strong>Problem</strong>:  Samples of known concentration vary consistently from the NUCLEIC dotMETRIC™ scale measurements.   </p>
<p><strong>Solution</strong>:  Personal technique and pipette tip size are the most common reasons for inaccurate measurements.  Measuring each sample at different dilutions in dilution buffer several times will normally provide very accurate results.  The NUCLEIC dotMETRIC™ scale was calibrated with fixed volume micro capillary tips, using the application device.  Use of other tips may influence dot diameter. Use Spot Application Device for better results.   </p>
<p> </p>

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