Difference between revisions of "Team:TP-CC San Diego/LabBook"

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<center><a href="https://static.igem.org/mediawiki/2017/8/81/T--TP-CC_San_Diego--labnotebook.pdf">Click Here for the full notebook PDF.</a></center>
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<center><a href="https://static.igem.org/mediawiki/2017/8/81/T--TP-CC_San_Diego--labnotebook.pdf">Click Here for the full notebook PDF</a></center>
 
     <h3 class = "day"> DAY 1 </h3>
 
     <h3 class = "day"> DAY 1 </h3>
 
     <div class = "description">  
 
     <div class = "description">  

Revision as of 04:54, 22 October 2017


Lab Notebook

Lab Notebook

Click Here for the full notebook PDF

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

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

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