Team:Stanford/Notebook/Lab Work/Week 1

From 2010.igem.org

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==Wednesday==
==Wednesday==
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''Laura'': Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed:
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get competent cells from -80oC freezer
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set H2O bath to 42oC (already done)
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add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
 +
add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time)
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on ice 30 min
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24oC H2O bath 20 sec.
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on ice 15 min
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add 1 mL SOC, 2 hours at 37oC (rotating)
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plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies
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variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal)
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Revision as of 22:47, 19 July 2010

Quad center.jpg

Spring: Brainstorming | Spring Meetings

Summer: Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Summaries


Contents

Monday

General Meeting Notes

  • Wiki stuff
    • Everyone should sign up on 2010.igem.org website
      • Take advantage of the new calendar system
      • Need to transfer current website over
    • Current wiki needs a new vision
      • Francisco + Alex assembling wiki committee
  • Transform the first wave of cells
    • Pick colonies
    • Colony PCR
    • Sequence parts to validate
      • Use the Smolke sequencing network
  • Characterize promoters
    • Digest, ligate
    • Need to acquire enzymes
    • Need an PO account.
      • Email Jeanne
  • Get competent cells
  • Getting DNA
    • Assembly PCR
    • PCR from host cells
    • Synthesize de novo
  • RNA team
    • Use RNAstructure
      • Maybe use ViennaRNA or dinamelt
    • Talk to Arkin and Berkeley Postdocs
    • Finalize sRNA library
    • 9kb plasmid
  • Meeting scheudle
    • Monday morning is good for logistics
    • Group meetings for presentations
      • One person presents their work in the context of everything else
    • Journal club
    • Karina will set up a doodle

Tuesday

Laura: made LB agar (with Alex and Chris)

  • made 1 500 mL bottle, 3 250 mL bottles, then autoclaved

Wednesday

Laura: Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed: get competent cells from -80oC freezer set H2O bath to 42oC (already done) add 50 uL cells to a 1.5 mL tube (keep tubes on ice) add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time) on ice 30 min 24oC H2O bath 20 sec. on ice 15 min add 1 mL SOC, 2 hours at 37oC (rotating) plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal)