Bitan:Scintillation counting using the Triathler bench-top counter

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 <p class=MsoNormal align=center style='text-align:center'><span
 style='font-family:Calibri'>Scintillation counting of labeled RNA<o:p></o:p></span></p>
 <ol style='margin-top:0cm' start=1 type=1>
  <li class=MsoNormal style='mso-list:l2 level1 lfo2;tab-stops:list 36.0pt'><span
      style='font-family:Calibri'>After </span><span style='font-family:Calibri;
      mso-fareast-language:KO'><i>in vitro</i></span><span style='font-size:
      16.0pt;font-family:Calibri'> </span><span style='font-family:Calibri'>transcription
      and solubilization of RNA product in 100 &#956;l (see <i><a
      href="http://openwetware.org/wiki/Bitan:In-vitro_transcription%2C_labeling_and_G-50_purification_of_RNA">In
      vitro<span style='font-style:normal'> transcription and RNA labeling</span></a></i></span><span
      style='font-family:Calibri'>) of the G-50 buffer, centrifuge the tube
      and keep 1 &#956;l of RNA product in a 0.6-ml tube for scintillation
      counting. <o:p></o:p></span></li>

  <li class=MsoNormal style='mso-list:l2 level1 lfo2;tab-stops:list 36.0pt'><span
      style='font-family:Calibri'>Perform counting using the Triathler
      bench-top scintillation counter (see below) and keep the aliquot for <a
      href="http://openwetware.org/index.php?title=Bitan:TBE-urea-polyacrylamide_gel_electrophoresis&amp;action=edit&amp;redlink=1">electrophoresis</a>.<o:p></o:p></span></li>
  <li class=MsoNormal style='mso-list:l2 level1 lfo2;tab-stops:list 36.0pt'><span
      style='font-family:Calibri'>Keep another 1-&#956;l aliquot of the RNA
      product after G-50 purification and perform counting. Keep the aliquot
      for <a
      href="http://openwetware.org/index.php?title=Bitan:TBE-urea-polyacrylamide_gel_electrophoresis&amp;action=edit&amp;redlink=1">electrophoresis</a>.<o:p></o:p></span></li>
  <li class=MsoNormal style='mso-list:l2 level1 lfo2;tab-stops:list 36.0pt'><span
      style='font-family:Calibri'>To perform counting <o:p></o:p></span></li>
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   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Start up the machine.<o:p></o:p></span></li>

   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Machine reads “Clear Saved Parameters”,
       “Powering up .. Version 1.8” and then either “&lt;H-3&gt; Ready start
       sys edit” or “P-32 Ready start sys edit”. <o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>If it does not read the latter, press
       “P-32” on the keyboard panel of the Triathler.<o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Insert the provided plastic scintillation
       adaptor into the counting chamber. This allows measurement of low-volume,
       high-energy, dry counting of the <sup>32</sup>P-labeled RNA samples.
       This also precludes the need to mix the sample with the scintillation
       fluid.<o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Insert the 0.6-ml tube with the 1-&#956;l
       aliquot of RNA inside the plastic adaptor inside the counting chamber;
       close the lid.<o:p></o:p></span></li>

   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Press start.<o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>If counting many samples, remove the first
       tube, insert the second tube and press next.<o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Jot down the counting values in
       cpm/&#956;l.<o:p></o:p></span></li>
   <li class=MsoNormal style='mso-list:l2 level2 lfo2;tab-stops:list 72.0pt'><span
       style='font-family:Calibri'>Divide the counting after G-50 purification
       to counting before purification to obtain percentage of incorporation
       of nucleotides (because G-50 removes unincorporated nucleotides).<o:p></o:p></span></li>
  </ol>
 </ol>

 <p class=MsoNormal align=center style='text-align:center'><span
 style='font-size:10.0pt;font-family:Calibri'><a
 href="http://openwetware.org/wiki/Bitan:todo">Back to To-Do List</a></span><span
 style='font-family:Calibri'><o:p></o:p></span></p>
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