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Revision as of 00:15, 30 October 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
