Team:Stanford/Notebook/Lab Work/Week 1

From 2010.igem.org

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===Francisco's Notebook===
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*Learned how to electroporate.
*helped Karina, Francisco with the transformations of parts
*helped Karina, Francisco with the transformations of parts

Revision as of 19:15, 27 July 2010

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Contents

6/28 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

6/29 Tuesday

Laura's Notebook

  • made LB agar (with Alex and Chris)
    • made 1 500 mL bottle, 3 250 mL bottles, then autoclaved

6/30 Wednesday

Laura's Notebook

  • Re-try failed transformation (failed 2X for Alex and Chris)
  • Protocol followed:
  1. get competent cells from -80oC freezer
  2. set H2O bath to 42oC (already done)
  3. add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
  4. add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time)
  5. on ice 30 min
  6. 24oC H2O bath 20 sec.
  7. on ice 15 min
  8. add 1 mL SOC, 2 hours at 37oC (rotating)
  9. 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)

7/1 Thursday

Laura's Notebook

  • did 8 minipreps from Chris'/Alex's cultures
  • used Qiagen spin kit, with the following variations:
  1. skipped "recommended" wash step
  2. eluted with H2O
  • Concentrations from Nanodrop (taken by Chris B.)
psb1A2: 58 ng/uL
psb1A3: 53 ng/uL
psb2K3: 40 ng/uL
psb4A5: 63.4 ng/uL
E0240: 65 ng/uL
T9002: 104 ng/uL
F2620: 57.5ng/uL

Francisco's Notebook

  • Learned how to electroporate.
  • helped Karina, Francisco with the transformations of parts
Retrieved from "http://2010.igem.org/Team:Stanford/Notebook/Lab_Work/Week_1"