Team:Illinois/Antibody GPCR Fusion Notebook

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!-- == 22nd July ==

  • Yeast obtained from Dr. Zhao


-- == 24th July ==

  • Prepared liquid culture for DNA extraction
  • Made 1M Tris. Cl pH 8.0
  • Made 4M ammonium acetate


!-- == 22nd August ==

  • Attempted DNA extraction
    • Result: Failed
  • Obtained more yeast from Dr. Zhao


!-- == 25th August ==

  • Prepared overnight culture for DNA extraction (3:27pm)


!-- == 26th August ==

  • Attempted DNA extraction
  • Prepped overnight culture


!-- == 27th August ==

  • Performed PCR (FILL IN PCR TABLE)
  • Prepped 3 overnight cultures


!-- == 28th August ==

  • Extracted DNA from 4 cultures
  • Ran gel of PCR products (1.5%, 200V)
    • Result: No bands present


!-- == 2nd September ==

  • FILL IN PCR TABLE


!-- == 3rd September ==

  • Ran gel
    • Ladder lane 7
    • Sample 7 spilled
    • 1% agarose
      • Too high
    • 120V
      • Too low
    • 50 minutes


!-- == 8th September ==

  • FILL IN PCR TABLE
  • Prepped 4 overnight cultures
    • Yeast dried out again


!-- == 9th September ==

  • Signs of life in 3 of the cultures
    • Wait until tomorrow
  • Ran gel on PCR from 8th September
    • 150V, 50 minutes
    • No sign of DNA
  • Ladder from Courtney


!-- == 10th September ==

  • Ran gel again
  • Split culture
    • 150V, 50 minutes
    • 0.75% gel
    • Ladder from Courtney


!-- == 11th September ==

  • FILL IN PCR TABLE


!-- == 12th September ==

  • Isolated DNA from 8 cultures
  • Ran gel
    • 1% agarose
    • 150V
    • 38 mins
      • Poor results


!-- == 15th September ==

  • PCR PGK Promotor
    • Finnzymes Phusion High Fidelity DNA Polymerase