Difference between revisions of "Team:Cardiff Wales/Notebook/Text"

Line 200: Line 200:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Ran colony PCR on new level 1 plates - gel wasn’t great (EH)</li>
 +
<li> PCR run with 14,15,16,18, primers 64+70 and 64+71 were used - bands for 64+71 were strong (ST and HE)</li>
 +
<li> Initial contact with WashU about potential collaboration via email (LT)</li>
 +
<li> Colony PCR on white colonies from the plates grown overnight. Ran the PCR products on a gel. Two colonies had bands around the expected size so were picked and grown overnight (EH) </li>
 +
<li> Replated the rest of the level 1 transformation mix onto new plates (EH)
 +
</li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 208: Line 213:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Repeated colony PCR on new level 1 plates - gel slightly better so put successful colonies to grow overnight (EH) </li>
 +
<li>Made new chloro broth (EH) </li>
 +
<li>Grew up level 0 parts in correct broth (EH) </li>
 +
<li>PCR run with 14,15,16,18, primers 69+70 and 69+71 were used. Bands for 69+70 were strong (ST and HE) </li>
 +
<li>Video call with WashU detailing possible collaboration (all) </li>
 +
<li>Miniprepped the two colonies grown overnight (EH) </li>
 +
<li>Colony PCR on the white colonies from the level 1 plates (35S/GFP/terminators) and PCR on the miniprepped colonies (EH) </li>
 +
<li>Ran the PCR products on the gel- only the miniprepped construct containing the G7 terminator had a band of the expected size (EH) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 216: Line 228:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Sent the level 1 construct containing 35S long/GFP/G7 off for sequencing with primer 69 (EH)</li>
 +
<li>Picked more white colonies off the terminator level 1 plates for PCR (EH)</li>
 +
<li>Ran the PCR products on a gel - there were no bands of the expected size (EH)</li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 224: Line 238:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li>Sent the 35S  long promoter off for sequencing with primer 57 (EH)</li>
 +
<li>Set up a level 1 reaction using 35S short/Enhancer/GFP/Terminators (EH)</li>
 +
<li>Transformation of the level 1 reaction - plate out onto Kan/IPTG/XGAL (EH) </li>
 +
<li>Regrow the 35S long overnight (EH)</li>
 +
 
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 232: Line 250:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Colony PCR on the white colonies from the level 1 reactions from yesterday (EH)  </li>
 +
<li>Run PCR products on a gel - most of the bands look good (EH) </li>
 +
<li> Pick and grow the colonies overnight which had bands of the expected size on the gel (EH) </li>
 +
<li> Miniprepped the 35S long which was grown overnight (EH) </li>
 +
<li> Further discussion on logo design (EM and RC) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 240: Line 262:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Sent 35S long for sequencing with primer 68 (EH) </li>
 +
<li> Miniprepped the cultures grown overnight (EH) </li>
 +
<li> Sent one of each level 1 construct with different terminators off for sequencing (35S/enhancer/GFP/terminators) (EH)  </li>
 +
<li> Agro transformation with the level 1 constructs containing GFP and different terminators (EH) </li>
 +
<li> Plants monitored and watered (EM) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 248: Line 274:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Grow seed overnight for making competent cells (EH)</li>
 +
<li> Made up a solution of MgCl2 and autoclaved it (EH) </li>
 +
<li> Obtaining contact information for potential collaborations (EM) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 256: Line 284:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Made competent <i> E. coli </i> (EH)  </li>
 +
<li> Transform new competent cells with 18J to test them - plate onto chloramphenicol and grow on the bench over the weekend. (EH) </li>
 +
<li> Streaked out the agro plates and grow over the weekend at 28℃. </li>
 +
<li> Sequencing came back for the 35S long, turned out it was 35S short which explains why the level 1s were not working.  </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 264: Line 295:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Pick colonies off the agro plates and grow cultures overnight in LB broth, Kan and Rif. </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 272: Line 303:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Infiltrated plants with 35S-GUS and either NosT, G7, or 35S (EH) </li>
 +
<li> Re-potted plants (EM) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 280: Line 312:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Level 1 Ligation for (15)cRTBVp_BCR3_Full and (18) 35S_BCR3_Full (HE) </li>
 +
<li> Transformation of cells with Level 1s (HE)</li>
 +
<li>Grew up four 11 35s (HE)</li>
 +
 
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 288: Line 323:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Preparation for Operation Earth. (LT) </li>
 +
<li> Checked on progress of re-potted plants. (EM) </li>
 +
<li> Miniprep of 35s (HE) </li>
 +
<li> RNA Extraction from plants 14 and 16 (+ve and -ve) (ST) </li>
 +
<li> Colony PCR for Plates 2 and 5 (HE) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 296: Line 335:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> RT Reaction (-ve, 14, 16) (ST) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 304: Line 343:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> Tested new primers with 14 and 16 (ST) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>
Line 312: Line 351:
 
<p>
 
<p>
 
<ul type="circle">
 
<ul type="circle">
−
<li> Add text here </li>
+
<li> PCR with cDNA + gDNA (ST) </li>
 
</ul>
 
</ul>
 
<br><br><br><br><br>
 
<br><br><br><br><br>

Revision as of 19:39, 19 September 2018


09/07/2018

  • First day in the lab
  • Had safety induction and signed appropriate forms






10/07/2018

  • Add text here






11/07/2018

  • Add text here






12/07/2018

  • Add text here






13/07/2018

  • Add text here






16/07/2018

  • Add text here






17/07/2018

  • Add text here






18/07/2018

  • Add text here






19/07/2018

  • Add text here






20/07/2018

  • Add text here






23/07/2018

  • Add text here






24/07/2018

  • Add text here






25/07/2018

  • Add text here






26/07/2018

  • Add text here






27/07/2018

  • Add text here






30/07/2018

  • Add text here






31/07/2018

  • Add text here






01/08/2018

  • Add text here






02/08/2018

  • Add text here






03/08/2018

  • Add text here






06/08/2018

  • Add text here






07/08/2018

  • Add text here






08/08/2018

  • Ran colony PCR on new level 1 plates - gel wasn’t great (EH)
  • PCR run with 14,15,16,18, primers 64+70 and 64+71 were used - bands for 64+71 were strong (ST and HE)
  • Initial contact with WashU about potential collaboration via email (LT)
  • Colony PCR on white colonies from the plates grown overnight. Ran the PCR products on a gel. Two colonies had bands around the expected size so were picked and grown overnight (EH)
  • Replated the rest of the level 1 transformation mix onto new plates (EH)






09/08/2018

  • Repeated colony PCR on new level 1 plates - gel slightly better so put successful colonies to grow overnight (EH)
  • Made new chloro broth (EH)
  • Grew up level 0 parts in correct broth (EH)
  • PCR run with 14,15,16,18, primers 69+70 and 69+71 were used. Bands for 69+70 were strong (ST and HE)
  • Video call with WashU detailing possible collaboration (all)
  • Miniprepped the two colonies grown overnight (EH)
  • Colony PCR on the white colonies from the level 1 plates (35S/GFP/terminators) and PCR on the miniprepped colonies (EH)
  • Ran the PCR products on the gel- only the miniprepped construct containing the G7 terminator had a band of the expected size (EH)






10/08/2018

  • Sent the level 1 construct containing 35S long/GFP/G7 off for sequencing with primer 69 (EH)
  • Picked more white colonies off the terminator level 1 plates for PCR (EH)
  • Ran the PCR products on a gel - there were no bands of the expected size (EH)






13/08/2018

  • Sent the 35S long promoter off for sequencing with primer 57 (EH)
  • Set up a level 1 reaction using 35S short/Enhancer/GFP/Terminators (EH)
  • Transformation of the level 1 reaction - plate out onto Kan/IPTG/XGAL (EH)
  • Regrow the 35S long overnight (EH)






14/08/2018

  • Colony PCR on the white colonies from the level 1 reactions from yesterday (EH)
  • Run PCR products on a gel - most of the bands look good (EH)
  • Pick and grow the colonies overnight which had bands of the expected size on the gel (EH)
  • Miniprepped the 35S long which was grown overnight (EH)
  • Further discussion on logo design (EM and RC)






15/08/2018

  • Sent 35S long for sequencing with primer 68 (EH)
  • Miniprepped the cultures grown overnight (EH)
  • Sent one of each level 1 construct with different terminators off for sequencing (35S/enhancer/GFP/terminators) (EH)
  • Agro transformation with the level 1 constructs containing GFP and different terminators (EH)
  • Plants monitored and watered (EM)






16/08/2018

  • Grow seed overnight for making competent cells (EH)
  • Made up a solution of MgCl2 and autoclaved it (EH)
  • Obtaining contact information for potential collaborations (EM)






17/08/2018

  • Made competent E. coli (EH)
  • Transform new competent cells with 18J to test them - plate onto chloramphenicol and grow on the bench over the weekend. (EH)
  • Streaked out the agro plates and grow over the weekend at 28℃.
  • Sequencing came back for the 35S long, turned out it was 35S short which explains why the level 1s were not working.






20/08/2018

  • Pick colonies off the agro plates and grow cultures overnight in LB broth, Kan and Rif.






21/08/2018

  • Infiltrated plants with 35S-GUS and either NosT, G7, or 35S (EH)
  • Re-potted plants (EM)






22/08/2018

  • Level 1 Ligation for (15)cRTBVp_BCR3_Full and (18) 35S_BCR3_Full (HE)
  • Transformation of cells with Level 1s (HE)
  • Grew up four 11 35s (HE)






23/08/2018

  • Preparation for Operation Earth. (LT)
  • Checked on progress of re-potted plants. (EM)
  • Miniprep of 35s (HE)
  • RNA Extraction from plants 14 and 16 (+ve and -ve) (ST)
  • Colony PCR for Plates 2 and 5 (HE)






24/08/2018

  • RT Reaction (-ve, 14, 16) (ST)






28/08/2018

  • Tested new primers with 14 and 16 (ST)






29/08/2018

  • PCR with cDNA + gDNA (ST)






30/08/2018

  • Add text here






31/08/2018

  • Add text here







03/09/2018

  • Add text here






04/09/2018

  • Add text here






05/09/2018

  • Add text here






06/09/2018

  • Add text here






07/09/2018

  • Add text here






10/09/2018

  • Add text here






11/09/2018

  • Add text here






12/09/2018

  • Add text here






13/09/2018

  • Add text here






14/09/2018

  • Last official day so cleaned up the lab (all)






17/09/2018

  • Add text here






18/09/2018

  • Add text here






19/09/2018

  • Add text here






20/09/2018

  • Add text here






21/09/2018

  • Add text here






21/09/2018

  • Add text here






24/09/2018

  • Add text here






24/09/2018

  • Add text here






25/09/2018

  • Add text here






26/09/2018

  • Add text here






27/09/2018

  • Add text here






28/09/2018

  • Add text here







31/10/2018

  • 2spooky4me