Difference between revisions of "Team:NU Kazakhstan/Project Timeline"

Line 301: Line 301:
 
</li>
 
</li>
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">11 June 2018</a>
+
<a href="#" class="float-right">13 June 2018</a>
 +
<p>Precipitate was detected in inoculated LB liquid medium.</p>
 +
<p>LB liquid medium with spectinomycin was prepared.</p>
 
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
 
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
 
</li>
 
</li>
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">11 June 2018</a>
+
<a href="#" class="float-right">14 June 2018</a>
−
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
+
<p>MiraPrep was done. </p>
 +
<p>Extracted plasmid samples were measured by Nanodrop.</p>
 +
<table>
 +
<tr>
 +
<td>Samples</td>
 +
<td>260/280</td>
 +
<td>260/230</td>
 +
<td>Concentration</td>
 +
</tr>
 +
<tr>
 +
<td>1</td>
 +
<td>1.85</td>
 +
<td>2.25</td>
 +
<td>1915 ng/ul</td>
 +
</tr>
 +
</table>
 +
<p>Gel Electrophoresis was done.</p>
 
</li>
 
</li>
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">11 June 2018</a>
+
<a href="#" class="float-right">15 June 2018</a>
−
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
+
<p>OD of liquid cyanobacteria culture was measured. OD: 1.035.</p>
 
</li>
 
</li>
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">11 June 2018</a>
+
<a href="#" class="float-right">16 June 2018</a>
−
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
+
<p>OD of liquid cyanobacteria culture was measured. OD: 1.5.</p>
 +
<p>Transformation of cyanobacteria (16/06/18, OD: 1.5) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 
</li>
 
</li>
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">11 June 2018</a>
+
<a href="#" class="float-right">17 June 2018</a>
−
<p>Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.</p>
+
<p>Transformed cyanobacteria (16/06/18)  were plated onto BG-11 agar plates with spectinomycin.</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week8"><u>WEEK 8 (June 18 – June 24)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">18 June 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>Extracted plasmid sample was linearized by digestion with EcoRI. </p>
 +
<p>Gel Electrophoresis done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">19 June 2018</a>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">20 June 2018</a>
 +
<p>Transformed cyanobacteria (19/06/18) cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>BG-11 medium was prepared.</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week9"><u>WEEK 9 (June 25 – July 1)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">25 June 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>BG-11 agar plates with spectinomycin were prepared.</p>
 +
<p>OD of liquid cyanobacteria culture was measured. OD: 1.5.</p>
 +
<p>Transformation of cyanobacteria (05/06/18, OD: 1.5) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">26 June 2018</a>
 +
<p>Transformed cyanobacteria (25/06/18)  were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">27 June 2018</a>
 +
<p>Transformed cyanobacteria (26/06/18) cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>OD of liquid cyanobacteria culture was measured. OD: 1.680.</p>
 +
<p>Transformation of cyanobacteria (05/06/18, OD: 1.680) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done. </p>
 +
<p>New subculture of cyanobacteria was prepared from old liquid culture (05/06/18, OD: 1.680)</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">28 June 2018</a>
 +
<p>Transformed cyanobacteria (27/06/18) cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>New subculture of cyanobacteria was prepared from old liquid culture (05/06/18).</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week10"><u>WEEK 10 (July 2 – July 8)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">2 July 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>New subculture of cyanobacteria was prepared from old liquid culture (20/06/18).</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">3 July 2018</a>
 +
<p>LB liquid medium and LB agar plates were prepared.</p>
 +
<p>BG-11 medium was prepared.</p>
 +
<p>New subculture of cyanobacteria was prepared from old liquid culture (20/06/18).</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">4 July 2018</a>
 +
<p>BG-11 agar plates with spectinomycin were prepared.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">5 July 2018</a>
 +
<p>Two transformations of cyanobacteria (OD: 0.511 and OD: 0.948) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) were done. </p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">6 July 2018</a>
 +
<p>Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>Colony segregation was done (colony from  27/05/18 was streaked onto new plate).</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week11"><u>WEEK 11 (July 9 – July 15)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">9 July 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>PCR amplification of SQR genes (Lep and Gei) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">10 July 2018</a>
 +
<p>OD of liquid cyanobacteria culture (20/06/18) was measured. OD: 1.562.</p>
 +
<p>Gel Electrophoresis with PCR products and Genes as control was run.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">11 July 2018</a>
 +
<p>Gel Electrophoresis with PCR products and Genes as control was run.</p>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">12 July 2018</a>
 +
<p>Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>PCR purification was done for PRC products by using PCR purification kit. Concentration of SQR Geitlerinema (SQR Gei): 91.23, SQR Leptolyngbya (SQR Lep): 81.66, SuperNova: 50.59 ng/ul. </p>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">13 July 2018</a>
 +
<p>Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week12"><u>WEEK 12 (July 16 – July 22)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">17 July 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>Interlab was started. </p>
 +
<p>OD of liquid cyanobacteria culture (02/07/18) was measured. OD: 0.8.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">18 July 2018</a>
 +
<p>PCR amplification of SQR Gei was done again and no favorable results were obtained.</p>
 +
<p>Gel Electrophoresis with PCR products was run.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">19 July 2018</a>
 +
<p>PCR amplification with restriction sites of  Lep, Lep with signal sequence (ss), Gei and Gei with signal sequence (ss).</p>
 +
<p>PCR purification was done for PRC products by using PCR purification kit.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">20 July 2018</a>
 +
<p>Lep, Lep with ss, Gei, Gei with ss and plasmid pSyn_6 were digested with BlII and KpnI restriction enzymes. </p>
 +
<p>Gel extraction was done.</p>
 +
<p>PCR amplification of Lep, Lep with ss, Gei and Gei with ss was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">21 July 2018</a>
 +
<p>Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 vector was done.</p>
 +
<p>Transformation of E.coli (TOP10) with modified plasmid pSyn_6 (SQR genes and marker gene: spectinomycin resistance gene) was done.  </p>
 +
<p>Transformed E.coli cells were plated onto LB agar plates with spectinomycin. </p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">22 July 2018</a>
 +
<p>Inoculation of the LB liquid medium with spectinomycin of resistant TOP10 cells was done.</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week13"><u>WEEK 13 (July 23 – July 29)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">23 July 2018</a>
−
<p>OD of liquid cyanobacteria culture was measured</p>
+
<p>Only transformed Gei E.coli grew up. </p>
 +
<p>MiraPrep was done. </p>
 +
<p>Extracted plasmid samples were measured by Nanodrop.</p>
 +
<table>
 +
<tr>
 +
<td>Samples</td>
 +
<td>260/280</td>
 +
<td>260/230</td>
 +
<td>Concentration</td>
 +
</tr>
 +
<tr>
 +
<td>Gei</td>
 +
<td>1.98</td>
 +
<td>2.35</td>
 +
<td>410.5 ng/ul</td>
 +
</tr>
 +
</table>
 +
<p>Gel Electrophoresis was done. No bands.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">24 July 2018</a>
 +
<p>Competent E.coli (TOP10) cells were prepared.</p>
 +
<p>OD of liquid cyanobacteria culture was measured.</p>
 +
<p>Untransformed E.coli was plated on a plate with spectinomycin to check antibiotic activity.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">25 July 2018</a>
 +
<p>Untransformed E.coli grew up in plate with spectinomycin. It was deduced that antibiotic in plates was degraded. </p>
 +
<p>Interlab was done. </p>
 +
<p>PCR amplification of SQR genes with different annealing temperature (Lep and Gei) was done. </p>
 +
<p>PCR purification was done for PRC products by using PCR purification kit.</p>
 +
<p>PCR products were measured by Nanodrop. </p>
 +
<table>
 +
<tr>
 +
<td>Samples</td>
 +
<td>260/280</td>
 +
<td>260/230</td>
 +
<td>Concentration</td>
 +
</tr>
 +
<tr>
 +
<td>Lep (72C)</td>
 +
<td>1.95</td>
 +
<td>1.48</td>
 +
<td>87.22 ng/ul</td>
 +
</tr>
 +
<tr>
 +
<td>Lep (80C)</td>
 +
<td>1.96</td>
 +
<td>3.04</td>
 +
<td>41.29 ng/ul</td>
 +
</tr>
 +
<tr>
 +
<td>Gei (72C)</td>
 +
<td>1.90</td>
 +
<td>1.39</td>
 +
<td>54.68 ng/ul</td>
 +
</tr>
 +
<tr>
 +
<td>Gei (80C)</td>
 +
<td>1.89</td>
 +
<td>1.19</td>
 +
<td>68.95 ng/ul</td>
 +
</tr>
 +
</table>
 +
<p>Lep, Lep with ss, Gei, Gei with ss and pSyn_6 plasmid were digested with BlII and KpnI restriction enzymes.</p>
 +
<p>Gel extraction was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">27 July 2018</a>
 +
<p>The ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done (1:10, insert : gene). Overnight incubation.</p>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">28 July 2018</a>
 +
<p>Transformed cyanobacteria (27/07/18) cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>Gel Electrophoresis of ligated pSyn_6 and SQR genes was run. SQR genes were not inserted.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">23 July 2018</a>
 +
<p>Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done (1:2, insert : gene). 2 hours incubation.</p>
 
</li>
 
</li>
 
</ul><br>
 
</ul><br>
 
<br>
 
<br>
 
 
−
<h4><b id="week4"><u>WEEK 5</u></b></h4>
+
<h4><b id="week4"><u>WEEK 14 (July 30 – August 5)</u></b></h4>
 
<ul class="timeline">
 
<ul class="timeline">
 
<li class="slideInRight wow">
 
<li class="slideInRight wow">
−
<a href="#" class="float-right">25 May 2018</a>
+
<a href="#" class="float-right">31 July 2018</a>
 +
<p>Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done. Stony Brook University protocol.</p>
 +
<p>New subculture of cyanobacteria was prepared from old liquid culture.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">1 August 2018</a>
 +
<p>Transformed cyanobacteria (31/06/18) cells were plated onto BG-11 agar plates with spectinomycin.</p>
 +
<p>PCR amplification of SQR genes was done.</p>
 +
<p>Gel Electrophoresis of PCR products was run.</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">3 August 2018</a>
 +
<p>Lep, Lep with ss, Gei, Gei with ss and plasmid pSyn_6 were digested with BlII and ScaI restriction enzymes.</p>
 +
<p>Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done.</p>
 +
<p>Transformation of E.coli (TOP10) with modified plasmid pSyn_6 (SQR genes and marker gene: spectinomycin resistance gene) was done.</p>
 +
<p>Transformed E.coli cells were plated onto LB agar plates with spectinomycin. </p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">4 August 2018</a>
 +
<p>OD of liquid cyanobacteria culture was measured</p>
 +
</li>
 +
<li class="slideInRight wow">
 +
<a href="#" class="float-right">5 August 2018</a>
 
<p>OD of liquid cyanobacteria culture was measured</p>
 
<p>OD of liquid cyanobacteria culture was measured</p>
 
</li>
 
</li>
Line 498: Line 713:
 
<h6 class="text-uppercase mb-20">Quick About</h6>
 
<h6 class="text-uppercase mb-20">Quick About</h6>
 
<p>
 
<p>
−
SCHOOL OF SCIENCE AND TECHNOLOGY <br> Nazarbayev University <br> Astana, Kazakhstan
+
SCHOOL OF SCIENCE AND TECHNOLOGY Nazarbayev University Astana, Kazakhstan
 
</p>
 
</p>
 
</div>
 
</div>

Revision as of 20:54, 17 October 2018

Bioremediation of Sour Crude Oil Waste using Cyanobacteria




May
Week 1-4
Week 1 April 30 - May 6
Week 2 May 7 - May 13
Week 3 May 14 - May 20
Week 4 May 21 - May 27
Week 5 May 28 – June 3
Week 6 June 4 – June 10
Week 7 June 11 – June 17
Week 8 June 18 – June 24
Week 9 June 25 – July 1

WEEK 1 (April 30 - May 6)

  • 5 May 2018

    BG-11 medium was prepared.

    Inoculation of the BG-11 medium with cyanobacteria (Synechococcus elongatus PCC 7942) was done.



WEEK 2 (May 7 - May 13)

  • 11 May 2018

    BG-11 agar plates were made.

    Cyanobacteria were plated onto BG-11 agar plates from liquid culture



WEEK 3 (May 14 - May 20)

  • 14 May 2018

    The new subculture of cyanobacteria was prepared from old liquid culture.

    The old liquid culture of cyanobacteria was plated onto BG -11 agar plate.

  • 16 May 2018

    BG-11 agar plates were prepared.

    Cyanobacteria from liquid culture were plated onto the BG-11 agar plates.



WEEK 4 (May 21 - May 27)

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured

    Liquid cultures of cyanobacteria were checked.

    The new subculture of cyanobacteria was prepared from old liquid culture.



WEEK 5 (May 28 – June 3)

  • 28 May 2018

    OD of liquid cyanobacteria culture was measured.

    Liquid cultures of cyanobacteria were checked.

  • 30 May 2018

    LB liquid medium and LB agar plates were prepared.

  • 31 May 2018

    Competent E.coli (TOP10) cells were prepared.



WEEK 6 (June 4 – June 10)

  • 4 June 2018

    New competent TOP10 cells were prepared.

    Spectinomycin stock solution 1000X was prepared.

    LB agar plates with spectinomycin were prepared.

  • 5 June 2018

    Transformation of E.coli (TOP10) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

    Transformed E.coli cells were plated on LB agar plates with spectinomycin.

  • 6 June 2018

    LB liquid medium with spectinomycin was prepared.

    Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.

  • 7 June 2018

    Plasmids from the transformed TOP10 were extracted by MiraPrep.

    Extracted plasmid samples were measured by Nanodrop.

    Samples 260/280 260/230 Concentration
    1 1.76 1.53 63.96 ng/ul
    2 1.96 2.29 1953 ng/u

    Gel Electrophoresis was done.

  • 8 June 2018

    OD of liquid cyanobacteria culture was measured. OD: 0.8.

    BG-11 agar plates with spectinomycin were prepared.



WEEK 7 (June 11 – June 17)

  • 11 June 2018

    Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.

  • 12 June 2018

    MiraPrep was done.

    Extracted plasmid samples were measured by Nanodrop.

    Samples 260/280 260/230 Concentration
    1 1.90 2.38 1600 ng/ul

    Gel Electrophoresis was done.

    Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.

  • 13 June 2018

    Precipitate was detected in inoculated LB liquid medium.

    LB liquid medium with spectinomycin was prepared.

    Inoculation of the LB liquid medium with spectinomycin resistant TOP10 cells was done.

  • 14 June 2018

    MiraPrep was done.

    Extracted plasmid samples were measured by Nanodrop.

    Samples 260/280 260/230 Concentration
    1 1.85 2.25 1915 ng/ul

    Gel Electrophoresis was done.

  • 15 June 2018

    OD of liquid cyanobacteria culture was measured. OD: 1.035.

  • 16 June 2018

    OD of liquid cyanobacteria culture was measured. OD: 1.5.

    Transformation of cyanobacteria (16/06/18, OD: 1.5) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 17 June 2018

    Transformed cyanobacteria (16/06/18) were plated onto BG-11 agar plates with spectinomycin.



WEEK 8 (June 18 – June 24)

  • 18 June 2018

    Extracted plasmid sample was linearized by digestion with EcoRI.

    Gel Electrophoresis done.

  • 19 June 2018

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 20 June 2018

    Transformed cyanobacteria (19/06/18) cells were plated onto BG-11 agar plates with spectinomycin.

    BG-11 medium was prepared.



WEEK 9 (June 25 – July 1)

  • 25 June 2018

    BG-11 agar plates with spectinomycin were prepared.

    OD of liquid cyanobacteria culture was measured. OD: 1.5.

    Transformation of cyanobacteria (05/06/18, OD: 1.5) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 26 June 2018

    Transformed cyanobacteria (25/06/18) were plated onto BG-11 agar plates with spectinomycin.

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 27 June 2018

    Transformed cyanobacteria (26/06/18) cells were plated onto BG-11 agar plates with spectinomycin.

    OD of liquid cyanobacteria culture was measured. OD: 1.680.

    Transformation of cyanobacteria (05/06/18, OD: 1.680) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

    New subculture of cyanobacteria was prepared from old liquid culture (05/06/18, OD: 1.680)

  • 28 June 2018

    Transformed cyanobacteria (27/06/18) cells were plated onto BG-11 agar plates with spectinomycin.

    New subculture of cyanobacteria was prepared from old liquid culture (05/06/18).



WEEK 10 (July 2 – July 8)

  • 2 July 2018

    New subculture of cyanobacteria was prepared from old liquid culture (20/06/18).

  • 3 July 2018

    LB liquid medium and LB agar plates were prepared.

    BG-11 medium was prepared.

    New subculture of cyanobacteria was prepared from old liquid culture (20/06/18).

  • 4 July 2018

    BG-11 agar plates with spectinomycin were prepared.

  • 5 July 2018

    Two transformations of cyanobacteria (OD: 0.511 and OD: 0.948) with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) were done.

  • 6 July 2018

    Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin.

    Colony segregation was done (colony from 27/05/18 was streaked onto new plate).



WEEK 11 (July 9 – July 15)

  • 9 July 2018

    PCR amplification of SQR genes (Lep and Gei) was done.

  • 10 July 2018

    OD of liquid cyanobacteria culture (20/06/18) was measured. OD: 1.562.

    Gel Electrophoresis with PCR products and Genes as control was run.

  • 11 July 2018

    Gel Electrophoresis with PCR products and Genes as control was run.

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 12 July 2018

    Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin.

    PCR purification was done for PRC products by using PCR purification kit. Concentration of SQR Geitlerinema (SQR Gei): 91.23, SQR Leptolyngbya (SQR Lep): 81.66, SuperNova: 50.59 ng/ul.

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 13 July 2018

    Transformed cyanobacteria cells were plated onto BG-11 agar plates with spectinomycin



WEEK 12 (July 16 – July 22)

  • 17 July 2018

    Interlab was started.

    OD of liquid cyanobacteria culture (02/07/18) was measured. OD: 0.8.

  • 18 July 2018

    PCR amplification of SQR Gei was done again and no favorable results were obtained.

    Gel Electrophoresis with PCR products was run.

  • 19 July 2018

    PCR amplification with restriction sites of Lep, Lep with signal sequence (ss), Gei and Gei with signal sequence (ss).

    PCR purification was done for PRC products by using PCR purification kit.

  • 20 July 2018

    Lep, Lep with ss, Gei, Gei with ss and plasmid pSyn_6 were digested with BlII and KpnI restriction enzymes.

    Gel extraction was done.

    PCR amplification of Lep, Lep with ss, Gei and Gei with ss was done.

  • 21 July 2018

    Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 vector was done.

    Transformation of E.coli (TOP10) with modified plasmid pSyn_6 (SQR genes and marker gene: spectinomycin resistance gene) was done.

    Transformed E.coli cells were plated onto LB agar plates with spectinomycin.

  • 22 July 2018

    Inoculation of the LB liquid medium with spectinomycin of resistant TOP10 cells was done.



WEEK 13 (July 23 – July 29)

  • 23 July 2018

    Only transformed Gei E.coli grew up.

    MiraPrep was done.

    Extracted plasmid samples were measured by Nanodrop.

    Samples 260/280 260/230 Concentration
    Gei 1.98 2.35 410.5 ng/ul

    Gel Electrophoresis was done. No bands.

  • 24 July 2018

    Competent E.coli (TOP10) cells were prepared.

    OD of liquid cyanobacteria culture was measured.

    Untransformed E.coli was plated on a plate with spectinomycin to check antibiotic activity.

  • 25 July 2018

    Untransformed E.coli grew up in plate with spectinomycin. It was deduced that antibiotic in plates was degraded.

    Interlab was done.

    PCR amplification of SQR genes with different annealing temperature (Lep and Gei) was done.

    PCR purification was done for PRC products by using PCR purification kit.

    PCR products were measured by Nanodrop.

    Samples 260/280 260/230 Concentration
    Lep (72C) 1.95 1.48 87.22 ng/ul
    Lep (80C) 1.96 3.04 41.29 ng/ul
    Gei (72C) 1.90 1.39 54.68 ng/ul
    Gei (80C) 1.89 1.19 68.95 ng/ul

    Lep, Lep with ss, Gei, Gei with ss and pSyn_6 plasmid were digested with BlII and KpnI restriction enzymes.

    Gel extraction was done.

  • 27 July 2018

    The ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done (1:10, insert : gene). Overnight incubation.

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done.

  • 28 July 2018

    Transformed cyanobacteria (27/07/18) cells were plated onto BG-11 agar plates with spectinomycin.

    Gel Electrophoresis of ligated pSyn_6 and SQR genes was run. SQR genes were not inserted.

  • 23 July 2018

    Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done (1:2, insert : gene). 2 hours incubation.



WEEK 14 (July 30 – August 5)

  • 31 July 2018

    Transformation of cyanobacteria with empty pSyn_6 vector (marker gene: spectinomycin resistance gene) was done. Stony Brook University protocol.

    New subculture of cyanobacteria was prepared from old liquid culture.

  • 1 August 2018

    Transformed cyanobacteria (31/06/18) cells were plated onto BG-11 agar plates with spectinomycin.

    PCR amplification of SQR genes was done.

    Gel Electrophoresis of PCR products was run.

  • 3 August 2018

    Lep, Lep with ss, Gei, Gei with ss and plasmid pSyn_6 were digested with BlII and ScaI restriction enzymes.

    Ligation of SQR genes (Lep, Lep with ss, Gei and Gei with ss) with pSyn_6 plasmid was done.

    Transformation of E.coli (TOP10) with modified plasmid pSyn_6 (SQR genes and marker gene: spectinomycin resistance gene) was done.

    Transformed E.coli cells were plated onto LB agar plates with spectinomycin.

  • 4 August 2018

    OD of liquid cyanobacteria culture was measured

  • 5 August 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured



WEEK 5

  • 25 May 2018

    OD of liquid cyanobacteria culture was measured

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