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

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<h3 class = "day"> DAY 14 </h3>
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            <b>SUMMARY of August 21st: </b>
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            <li>seed ~1.2 x 107 293T cells in a 10 cm dish </li>
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<h3 class = "day"> DAY 15 </h3>
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            <b>SUMMARY of August 17th: </b>
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            <li>Prepared the transfection complex. </li>
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Revision as of 01:28, 2 November 2017

Lab Notebook

Click Here to Refer to Protocols Page

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.