Difference between revisions of "Team:Dundee/CGCraigon"

Line 175: Line 175:
 
       <div class="border-day">
 
       <div class="border-day">
 
         <div class="box">
 
         <div class="box">
−
             <span class="box-title">26/6: Preparation of overnight cultures of positive colonies of pSb1C3-<I>OBP2A</I> from the transformation performed on the 25/6.  </span>
+
             <span class="box-title">26/6: Preparation of overnight cultures of pSb1C3-<I>OBP2A</I> from the transformation performed on the 25/6.  </span>
 
               <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew2-5"></span>
 
               <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew2-5"></span>
 
                 <div id="nmcollapsiblew2-5" class="collapse box-content">
 
                 <div id="nmcollapsiblew2-5" class="collapse box-content">
Line 187: Line 187:
 
         </div>
 
         </div>
 
       </div>
 
       </div>
 +
<div class="row">
 +
      <div class="border-day">
 +
        <div class="box">
 +
            <span class="box-title">27/6: Plasmid preparation and pre-sequence digest of pSB1C3-<I>OBP2A</I>.  </span>
 +
              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew2-6"></span>
 +
                <div id="nmcollapsiblew2-6" class="collapse box-content">
 +
                  <p><b>Aim of experiment:</b> To perform a plasmid preparation of overnight cultures of pSB1C3-<i>OBP2A</i> from positive colonies of JM110 <I>E.coli</I> strain from the transformation done on the 25/6. Then to subsequently perform a pre-sequence digest of the plasmid preparation. 
 +
                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
 +
        <p><b>Results:</b> Insert table values!
 +
 +
 +
                  <p><b>Next Steps:</b> Looking at the pre-sequence digest it has been decided that sample 5 will be sent for sequencing. This will be done on the 29/6.  </div>
 +
        </div>
 +
        </div>
 +
      </div>
 +
  
  

Revision as of 23:30, 6 September 2015

CSI: DUNDEE

The Forensic Toolkit

Watch Our Introduction Video
Nasal Mucus
Week Beginning 15/6/2015

Summary

Produced a plasmid preperation of pIDT-OBP2A in order to use it as template in PCR.

17/6: Transformation of pIDT-OBP2A in MC1610 E.coli strain.

Aim of Experiment: To transform pIDT-OBP2A into MC1061 e.coli strain

Protocols Used: Transformation

Results: N/A

Next Steps: If the transformation is successful the next step will be to produce an overnight culture for plasmid preparation.

18/6: Produce an overnight culture of pIDT-OBP2A plasmid in MC1061.

Aim of experiment: To produce an overnight culture of pIDT-OBP2A from positive colonies of MC1061 E.coli strain from the transformation done on the 17/6.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Produce a plasmid preparation of pIDT-OBP2A from this overnight culture.

19/6: Produce a plasmid preparation of pIDT-OBP2A from the overnight culture.

Aim of experiment: The overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain the pIDT-OBP2A plasmid.

Protocols Used: Miniprep

Results: The plasmid preperation yeilded a concentration of 305.53 ng/ul.

Next Steps: To run a PCR using the purified pIDT-OBP2A plasmid as a template for amplification.

Week Beginning 22/6/2015

Summary

Removal of the signaling peptide from OBP2A and insertion of OBP2A into the biobrick vector pSB1C3.

22/6: PCR of pIDT-OBP2A using primers designed for the removal of the signaling peptide.

Aim of Experiment: To perform a PCR on the plasmid preparation of pIDT-OBP2A produced last week, using primers that have been designed in a way as to remove the signalling peptide at the start of the OBP2A gene.

Protocols Used: PCR

Results: N/A

Next Steps: The next step will be to run the PCR product on a gel and perform a gel extraction on the part of the gel corresponding to the size of the OBP2A gene fragment.

23/6: Gel extraction of OBP2A from the PCR performed on the 22/6.

Aim of experiment: To perform a gel extraction of OBP2A from a PCR reaction mixture that was performed on the 22/6. Once gel extraction has been performed a subsequent gel will be done to determine if the gel extraction has been successful.

Protocols Used: Gel extraction

Results: As it can be seen from the gel image, a band is present corresponding to the size of the OBP2A gene fragment.

Next Steps: To perform a restriction of OBP2A in preparation for ligation.

24/6: Restriction, ligation and transformation of OBP2A into pSB1C3.

Aim of experiment: To firstly restrict the gel extracted OBP2A. Secondly, to then ligate the restricted OBP2A into the biobrick vector pSB1C3. Third and finally to transform this plasmid into the JM110 E.coli strain.

Protocols Used: Restriction Digest : Ligation : Transformation

Results: N/A

Next Steps:If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep.

25/6: Religation and transformation of OBP2A into pSB1C3.

Aim of experiment: The transformation performed on the 24/6 failed and as a result OBP2A will again ligated into the biobrick vector pSB1C3 and then transformed into JM110 E.coli strain.

Protocols Used: Ligation: Transformation

Results: N/A

Next Steps:If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep.

26/6: Preparation of overnight cultures of pSb1C3-OBP2A from the transformation performed on the 25/6.

Aim of experiment: To produce overnight cultures of pSB1C3-OBP2A from positive colonies of JM110 E.coli strain from the transformation done on the 25/6.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Produce a plasmid preparation of pSB1C3-OBP2A in preparation for pre-sequence digest.

27/6: Plasmid preparation and pre-sequence digest of pSB1C3-OBP2A.

Aim of experiment: To perform a plasmid preparation of overnight cultures of pSB1C3-OBP2A from positive colonies of JM110 E.coli strain from the transformation done on the 25/6. Then to subsequently perform a pre-sequence digest of the plasmid preparation.

Protocols Used: Overnight Cultures

Results: Insert table values!

Next Steps: Looking at the pre-sequence digest it has been decided that sample 5 will be sent for sequencing. This will be done on the 29/6.