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[DESCRIPTION]
<p>Measurement of DNA can be done using two broad methods (Spectrophotometric or Fluorometric).</p>
<p> </p>
<p>Qubit is a fluorometric method and uses a double stranded DNA specific dye. Therefore the sensitivity is higher and impurities like RNA or nucleotides in the sample will not affect the quantification as it would in spectrophotometric methods like a Nanodrop.<br /><br />The Qubit is first calibrated using two standards. To make a measurement, 199ul of buffer +1ul of dye + 1ul of sample is added to a qubit tube, which is inserted into the instrument.<br /><br />Here, we use OpenTrons to prepare samples for qubit measurement. It also saves time by requiring only one initial calibration for each container. It uses p1000 to distribute master mix and p10 to distribute unique samples.</p>
<p> </p>

[GUIDELINES]
<p><strong><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/rvpgazn.jpg" width="796" height="447" data-src="https://s3.amazonaws.com/pr-journal/rvngazn.jpg" data-ofn="Slide1.jpg" /></strong></p>
<p> </p>
<p><a href="https://s3.amazonaws.com/pr-journal/rvpgazn.jpg" target="_blank">Enlarge in New Window</a></p>
<p> </p>
<p><strong>Brief Overview:</strong></p>
<p><br />Measurement of DNA can be done using two broad methods (Spectrophotometric or Fluorometric).</p>
<p> </p>
<p>Qubit is a fluorometric method and uses a double stranded DNA specific dye. Therefore the sensitivity is higher and impurities like RNA or nucleotides in the sample will not affect the quantification as it would in spectrophotometric methods like a Nanodrop.<br /><br />The Qubit is first calibrated using two standards. To make a measurement, 199ul of buffer +1ul of dye + 1ul of sample is added to a qubit tube, which is inserted into the instrument.<br /><br />Here, we use OpenTrons to prepare samples for qubit measurement. It also saves time by requiring only one initial calibration for each container. It uses p1000 to distribute master mix and p10 to distribute unique samples.</p>
<p> </p>
<p>Please see the flowchart of the protocol below.</p>
<p> </p>
<p><strong>Materials:</strong></p>
<p> </p>
<p>Qubit dsDNA HS Assay Kit (1)</p>
<p>Invitrogen catalog number: Q32851</p>
<p> </p>
<p>Qubit Assay Tube (1 per sample)</p>
<p>Invitrogen catalog number: Q32856</p>
<p> </p>
<p>Run Samples Placed in Strip Tubes</p>
<p> </p>
<p><strong>Instruments:</strong></p>
<p> </p>
<p>Qubit Fluorometer (1)</p>
<p>Invitrogen catalog number: Q32866</p>
<p> </p>
<p>14ml Falcon Tube (1)</p>
<p>Alkali Scientific catalog number: CT5250</p>
<p> </p>
<p>14ml Tube Rack (1)</p>
<p> </p>
<p>96-well Qubit Tube Rack (1-2)</p>
<p> </p>
<p>96-well Strip Tube Rack (1)</p>
<p> </p>
<p>1000µl Tips</p>
<p>Fisher catalog number: 13-611-126</p>
<p> </p>
<p>10µl Tips</p>
<p>Eppendorf catalog number: 022491504</p>
<p> </p>
<p>OpenTrons Pro (1)</p>
<p> </p>
<p>Racks to hold tubes (Pictured in steps section)</p>
<p> </p>
<p> </p>
<p> </p>
<p>Link to download graphviz code to generate this flowchart:</p>
<p><a href="https://www.protocols.io/download?s3url=https://s3.amazonaws.com/pr-journal/rrzgazn.dot&amp;ofn=qubit.dot" target="_blank">Graphviz Dot File</a></p>
<p> </p>
<p><img src="https://s3.amazonaws.com/pr-journal/rssgazn.png" data-src="https://s3.amazonaws.com/pr-journal/rsrgazn.png" data-ofn="flowchart.png" /></p>

[STEPS]
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