Team:UIUC-Illinois/Notebook/LabNotebook

Week 1 Notes

5/29/2012	- Designed 9 primers for LacI. LacZ. and eYFP. Prepared LB. - Inoculation of B0014.J23100.wtPUF. B0030.Q04121 - Transformed the psB1C3 biobrick into DH5a cells
5/31/2012	- Transformed C0012, I732005, E0030, I732017. Also transformed PSB1A3, PSB1AK3, PSB1K3. - Inoculated LB media with a psB1C3 colony from the plate grown the day before. - Met with Prof. Bhalerao to talk about PUF application and project direction (RNA scaffolding).
6/1/2012	- Short-term goal changed: make one testing construct, [rbs-puf-eYFP] cloned into P.protet plasmid with a ptet promoter and terminator - Began inoculation of C0012, I732005, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3. Was incubated for 9 hours, then put in 4 degree room - Inoculated again due to the round-bottom tube being misplaced. Mini-prepped the inoculation.
6/2/2012	- Cultured [PUF+PIN], E0030, and P. protet. designed check primers for [rbs-puf-eYFP]
6/3/2012	- Inoculated with a psB1C3 colony in order to prepare a glycerol stock of the strain.

Week 2 Notes

6/4/2012	- Threw out old, incomplete inoculations from Friday. - Transformed I732005, R0010. - Miniprepped Venus and E0030 and put gDNA in the -20 temp box. - Did new inoculations on C0012, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3. - Made a glycerol stock of the psB1C3 strain.
6/5/2012	- Made glycerol stocks of C0012, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3 and put them in the -80 freezer. - Transformation of I732005 failed, so replated with rest of culture. - Transformed J04500 and PSB3T5. - Minipreped PUF Amp and protet CM20 (spilled 1 tube of this) and put gDNA in the -20 degree in the temp box. - Inoculated R0010.
6/7/2012	- Set goal to create RNA scaffold by 6/15, begun design.

6/11/2012	- Innoculated K12 - Transformed split GFP constructs BBa_K157005 and BBa_K157006 into DH5a cells - Constructed the first RNA scaffold draft - Inoculated both parts of split GFP, as well as J04500
6/12/2012	- Miniprepped K12 cells - RNA scaffold design will inculde two scaffolds which need to be stored into one plasmid and then ligated into separate plasmids - Made glycerol stock of both split GFP parts, J04500, and miniprepped both split GFP
6/13/2012	- Transformed Pertobrick into K12 cells - Constructed the first draft of the scaffold, but it needs review. - Second, unique scaffold needs to be designed
6/14/2012	- Plated Petrobrick transformations; inoculated cell growth using CM resistance - Finalized an updated version of the scaffold based on new data of PUF binding. Final modifications need to be made before sent for synthesis. - Performed PCR: (2) 50 uL tubes of control (no PUF binding site) and (2) 50 uL tubes with the PUF binding site. Used WT PUF as a template and phusion as the polymerase. Ran gel and saw a result of all primer dimers.
6/15/2012	- Made plan to digest cells in order to see whether or not petrobrick was transformed into K12. - Final modifications made on the scaffold design. The construct is ready for synthesis. - Second try of PCR: Did a temperature gradient of 55-63-71 degrees Celsius. Used the WT PUF as template DNA. Gel results show only primer dimers, the faintest of faint lines at around 700 base pairs, but it's not a positive result. - Miniprepped pTET is in the "temporary" box of the -20 degree Celsius freezer.

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@@ Line 104: / Line 104: @@
 </div>
-<div id="week3puf" style="display:none">
+<div id="week3puf" style="display:none">
+<table border=1>
+  <tbody>
+    <tr>
+      <td>6/11/2012</td>
+      <td>- Innoculated K12 <br />- Transformed split GFP constructs BBa_K157005 and BBa_K157006 into DH5a cells<br />- Constructed the first RNA scaffold draft<br />- Inoculated both parts of split GFP, as well as J04500</td>
+    </tr>
+    <tr>
+      <td><br />6/12/2012</td>
+      <td>- Miniprepped K12 cells<br />- RNA scaffold design will inculde two scaffolds which need to be stored into one plasmid and then ligated into separate plasmids<br />- Made glycerol stock of both split GFP parts, J04500, and miniprepped both split GFP</td>
+    </tr>
+    <tr>
+      <td>6/13/2012</td>
+      <td>- Transformed Pertobrick into K12 cells<br />- Constructed the first draft of the scaffold, but it needs review. <br />- Second, unique scaffold needs to be designed<br /></td>
+    </tr>
+    <tr>
+      <td>6/14/2012</td>
+      <td>- Plated Petrobrick transformations; inoculated cell growth using CM resistance<br />- Finalized an updated version of the scaffold based on new data of PUF binding. Final modifications need to be made before sent for synthesis. <br />- Performed PCR: (2) 50 uL tubes of control (no PUF binding site) and (2) 50 uL tubes with the PUF binding site. Used WT PUF as a template and phusion as the polymerase. Ran gel and saw a result of all primer dimers. <br /></td>
+    </tr>
+    <tr>
+      <td>6/15/2012</td>
+      <td>- Made plan to digest cells in order to see whether or not petrobrick was transformed into K12. <br />- Final modifications made on the scaffold design. The construct is ready for synthesis.<br />- Second try of PCR: Did a temperature gradient of 55-63-71 degrees Celsius. Used the WT PUF as template DNA. Gel results show only primer dimers, the faintest of faint lines at around 700 base pairs, but it's not a positive result.  <br />- Miniprepped pTET is in the "temporary" box of the -20 degree Celsius freezer.</td>
+    </tr>
+  </tbody>
+</table>
 </div>

Team:UIUC-Illinois/Notebook/LabNotebook

From 2012.igem.org

Revision as of 04:41, 3 October 2012

Lab Notebook

Week 1 Notes

Week 2 Notes