Difference between revisions of "Team:TP-CC San Diego/LabBook"
| Line 41: | Line 41: | ||
<div class = "text-wrapper"> | <div class = "text-wrapper"> | ||
| − | <center><a href="https://2017.igem.org/Team:TP-CC_San_Diego/Protocols | + | <center><a href="https://2017.igem.org/Team:TP-CC_San_Diego/Protocols">Click Here to Refer to Protocols Page</a></center> |
| − | ">Click Here to Refer to Protocols Page</a></center> | + | |
<h3 class = "day"> DAY 1 </h3> | <h3 class = "day"> DAY 1 </h3> | ||
<div class = "description"> | <div class = "description"> | ||
| Line 202: | Line 201: | ||
| − | + | ||
<h3 class = "day"> DAY 15 </h3> | <h3 class = "day"> DAY 15 </h3> | ||
| Line 214: | Line 213: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
<h3 class = "day"> DAY 16 </h3> | <h3 class = "day"> DAY 16 </h3> | ||
| Line 227: | Line 226: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
<h3 class = "day"> DAY 17 </h3> | <h3 class = "day"> DAY 17 </h3> | ||
| Line 239: | Line 238: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
<h3 class = "day"> DAY 18 </h3> | <h3 class = "day"> DAY 18 </h3> | ||
| Line 251: | Line 250: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
| Line 264: | Line 263: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | |||
<h3 class = "day"> DAY 20 </h3> | <h3 class = "day"> DAY 20 </h3> | ||
| Line 276: | Line 274: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
<h3 class = "day"> DAY 21 </h3> | <h3 class = "day"> DAY 21 </h3> | ||
| Line 289: | Line 287: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | + | ||
<h3 class = "day"> DAY 22 </h3> | <h3 class = "day"> DAY 22 </h3> | ||
| Line 328: | Line 326: | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | |||
| − | |||
| − | |||
| − | |||
</div> | </div> | ||
| − | |||
</body> | </body> | ||
</html> | </html> | ||
Revision as of 01:37, 2 November 2017
Lab Notebook
DAY 1
SUMMARY of July 17th:
- DNA extraction from GBM39 cells
- Agarose gel electrophoresis to confirm DNA
DAY 2
SUMMARY of July 18th:
- qPCR primer design and order
DAY 3
SUMMARY of July 21:
- Primers arrived and diluted according to instructions
DAY 4
SUMMARY of August 1st:
- qPCR for EGFR gene count and guide RNA design; placed order
DAY 5
SUMMARY of August 2nd:
- Received oligos and diluted according to instruction.
DAY 6
SUMMARY of August 7th:
- Digestion of lentiviral CRISPR plasmid
- Gel purification digested plasmid using QIAquick Gel Extraction Kit and eluted in buffer EB
- There should be two bands on the gel, one at 2kb which should not be purified. Purify only the larger band
DAY 7
SUMMARY of August 9th:
- Anneal oligo pairs
- Ligation and transformation
DAY 8
SUMMARY of August 10th:
- No colonies grew on carb antibiotic plates
DAY 9
SUMMARY of August 11th:
- Used same,remaining ligation reaction and transformed again onto carb antibiotic plates
DAY 10
SUMMARY of August 14th:
- Inoculate comp cells
DAY 11
SUMMARY of August 15th:
- Miniprep gRNA plasmids and sent in for sequencing
- Start interlab study
- Ludox Calibration
- Fluorescein Dilution
- Transform plasmids from plate 7 in kit provided by IGEM for interlab study
DAY 12
SUMMARY of August 16th:
- Received sequencing results and all plasmids have the gRNA
- Inoculate plasmids for interlab study
DAY 13
SUMMARY of August 17th:
- Interlab cell measurement
DAY 14
SUMMARY of August 21st:
- seed ~1.2 x 107 293T cells in a 10 cm dish
DAY 15
SUMMARY of August 17th:
- Prepared the transfection complex.
DAY 16
SUMMARY of August 23rd:
- Removed the transfection medium from cells
- Miniprep plasmids and measure concentration
DAY 17
SUMMARY of August 24th:
- Harvest cells
DAY 18
SUMMARY of August 25th:
- Plate target cells, incubate overnight
DAY 19
SUMMARY of August 26th:
- Remove the cell culture medium and replace with fresh medium, incubate overnight
DAY 20
SUMMARY of August 27th:
- Split cells 1:3 and incubate for 48 hours
DAY 21
SUMMARY of August 29th:
- Infected cells are selected for stable expression
- Continue monitoring cells
DAY 22
SUMMARY of October 21st:
- Digest guideRNA using restriction enzymes out of TLCV2 vector
- Run 1% agarose gel and cut out insert
- Gel purify insert and vector(pSB1C3) given by iGEM
- Ligation of guide RNA insert and pSB1C3 vector
- Transformation of plasmid for part submission
DAY 23
SUMMARY of October 22st:
- Inoculated plasmid for part submission
DAY 24
SUMMARY of October 23st:
- IMiniprep plasmids and measure concentration
