Team:Illinois/Antibody GPCR Fusion Notebook

From 2008.igem.org

(Difference between revisions)
Line 93: Line 93:
* PCR PGK Promotor
* PCR PGK Promotor
** Finnzymes Phusion High Fidelity DNA Polymerase
** Finnzymes Phusion High Fidelity DNA Polymerase
 +
*** F-530, 20V (2V/uL)
 +
 +
**FiLL out PCR TABLE
 +
 +
 +
== 18th September ==
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* Ran ___? reaction
 +
* Extracted DNA from gel from 8th September (PGK Terminator)
 +
*FILL TABLE
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 +
 +
== 19th September ==
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* Gel of PGK Promotor has no DNA present
 +
 +
 +
== 23rd September ==
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* PCR: Fus1 Downstream
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* Fill PCR TABLE
 +
 +
 +
== 24th September ==
 +
* Ran gel of Fus1 Downstream
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** Result: No DNA present on gel
 +
 +
 +
== 25th September ==
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* PCR: Fus1 Upstream
 +
 +
 +
== 1st October ==
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* PCR: Ste2
 +
 +
 +
== 8th October ==
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* PCR: PGK Terminator
 +
** Use DNA extracted from gel on 18th September
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* Also extracted DNA from gel from 30th September
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 +
 +
 +
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Revision as of 01:48, 10 October 2008

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Contents

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
      • F-530, 20V (2V/uL)
    • FiLL out PCR TABLE


18th September

  • Ran ___? reaction
  • Extracted DNA from gel from 8th September (PGK Terminator)
  • FILL TABLE


19th September

  • Gel of PGK Promotor has no DNA present


23rd September

  • PCR: Fus1 Downstream
  • Fill PCR TABLE


24th September

  • Ran gel of Fus1 Downstream
    • Result: No DNA present on gel


25th September

  • PCR: Fus1 Upstream


1st October

  • PCR: Ste2


8th October

  • PCR: PGK Terminator
    • Use DNA extracted from gel on 18th September
  • Also extracted DNA from gel from 30th September





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