Team:Stanford/Notebook/Lab Work/Week 1
From 2010.igem.org
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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
- Everyone should sign up on 2010.igem.org website
- 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
- Use RNAstructure
- 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 Section
made LB agar (with Alex and Chris)
- made 1 500 mL bottle, 3 250 mL bottles, then autoclaved
6/30 Wednesday
Laura's Notebook Section
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)