Bitan:Measuring DNA concentration by absorbance

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border:none;mso-border-alt:solid windowtext .5pt;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'>
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 <td width=443 valign=top style='width:442.8pt;border:solid windowtext .5pt;
 background:#666666;mso-shading:windowtext;mso-pattern:solid #666666;
 padding:0cm 5.4pt 0cm 5.4pt'>
 <p class=MsoNormal align=center style='text-align:center'><span
 style='font-size:14.0pt;font-family:Calibri;color:#CCFFFF'><b>Measuring DNA
 product by UV absorbance<o:p></o:p></b></span></p>
 <p class=MsoNormal><span style='font-family:Calibri;color:#CCFFFF'><![if !supportEmptyParas]>&nbsp;<![endif]><o:p></o:p></span></p>
 <p class=MsoNormal><span style='font-family:Calibri;color:#CCFFFF'>The
 Eppendorf BioPhotometer (in Dr. Spencer’s Lab.) was used.<span
 style="mso-spacerun: yes">&nbsp; </span><o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1;tab-stops:list 18.0pt'><![if !supportLists]><span
 style='font-family:Calibri;color:#CCFFFF'>1.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Switch
 ON.<o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1'><![if !supportLists]><span style='font-family:Calibri;
 color:#CCFFFF'>2.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Select
 dsDNA from the options, showing 1 OD<sub>260</sub> = 50 &#956;g/ml.<o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1'><![if !supportLists]><span style='font-family:Calibri;
 color:#CCFFFF'>3.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Add
 100 ml water to the plastic cuvette appropriate for use on this machine, and
 press ‘Blank’.<span style="mso-spacerun: yes">&nbsp; </span>The monitor
 should show ‘0.000’.<o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1'><![if !supportLists]><span style='font-family:Calibri;
 color:#CCFFFF'>4.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Add
 50 or 100-fold diluted DNA sample in the cuvette, insert the cuvette into the
 reader chamber, and press Sample.<span style="mso-spacerun: yes">&nbsp;
 </span>The minimum required for measurement is 50 &#956;l.<span
 style="mso-spacerun: yes">&nbsp; </span><o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1'><![if !supportLists]><span style='font-family:Calibri;
 color:#CCFFFF'>5.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Aspirate
 the sample off the cuvette and add the next sample. There is no need to wash
 between sample measurements.<o:p></o:p></span></p>
 <p class=MsoNormal style='margin-left:18.0pt;text-indent:-18.0pt;mso-list:
 l11 level1 lfo1'><![if !supportLists]><span style='font-family:Calibri;
 color:#CCFFFF'>6.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
 </span></span><![endif]><span style='font-family:Calibri;color:#CCFFFF'>Record
 the results. Calculate the actual concentration of purified DNA for the next </span><span
 style='font-family:Calibri;color:#CCFFFF;mso-fareast-language:KO'><i>in vitro</i></span><span
 style='font-size:16.0pt;font-family:Calibri;color:#CCFFFF'> </span><span
 style='font-family:Calibri;color:#CCFFFF'>transcription step.<span
 style="mso-spacerun: yes">&nbsp; </span>Take into account the dilution factor
 used. For reference, when A260/A280 is 1.8 for DNA or 2.0 for RNA, the sample
 is ultra-pure.<o:p></o:p></span></p>
 <p class=MsoNormal><span style='font-family:Calibri;color:#CCFFFF'>The same
 measurements can be performed in our lab using the old spectrophotometer and
 the Quartz cuvette. Make sure the cuvette is washed properly because the
 residual protein may lead to erroneous A260/A280 ratios.<o:p></o:p></span></p>
 <p class=MsoNormal align=center style='text-align:center'><span
 style='font-family:Calibri;color:#CCFFFF'><![if !supportEmptyParas]>&nbsp;<![endif]><o:p></o:p></span></p>
 <p class=MsoNormal align=center style='text-align:center'><span
 style='font-family:Calibri;color:#CCFFFF'><a
 href="http://openwetware.org/wiki/Bitan:todo"><span style='color:#CCFFFF'>Back
 to To-Do List</span></a></span><span style='font-family:Calibri;color:#33CCCC'><o:p></o:p></span></p>
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