Team:Brown-Stanford/Lab/Notebook/Week0

• REGObricks
  Intro --------------------- ISRU --------------------- Biocement --------------------- S. pasteurii --------------------- Balloon
  Flights --------------------- Transforming --------------------- Biobrick²
• PowerCell
  Intro --------------------- Cyanobacteria --------------------- Mars --------------------- Nutrients
• FRETcetera
  Intro --------------------- FRET etc. --------------------- Construct
• Lab
  Team --------------------- Parts --------------------- Notebook --------------------- Protocols --------------------- Safety
• Partners
  Collab --------------------- Sponsors
• Outreach
  SB5.0 --------------------- NASA --------------------- Lunar Science --------------------- Maker Faire --------------------- BBC --------------------- CBricks --------------------- Ethics --------------------- Regional
  Jamboree

June 20, 2011

• Made solid, liquid LB
• Made kan, amp solid LB plates
• made CCMB80 Buffer
• Made BG11 Plates
• Streaked out Golden strains on BG11 plates (in the 30? incubator)
• Created 5mL liquid BG11 cultures of all the Golden strains (in the 30? incubator)
• Streaked Norman’s E. coli conjugative plasmid strains on ampicilin to verify plasmid retention

June 21, 2011

Designed construct (see construct page)

Designed Co-culture experiment (see experiment design page)

Incubate with shaking (110rpm) at 30?

Max created liquid culture of Norman’s RP1 strain

June 22, 2011

Oops, shouldn’t have incubated with shaking. Without shaking until green, then move to shaking at 110rpm. Took cyano liquid cultures off shaking platform.

Created liquid culture of Silver’s sucrose invertase/glucose export S. elongatus strain in BG11 Created stocks for plasmid prep

June 23, 2011

Made BG11 stocks except Na2CO3 Made 50mg/mL streptomycin stock

Plasmid prepped Norman’s strains using Norman’s plasmid prep protocol:

• D1: pUC18@DH5a NW20110620.41
• D2: pSB1A3-P1010@DB3.1 NW20110620.A2
• D3: pSB1A10-P1010@DB3.1 NW20110620.A3
• D4: pSB1A10-J133452@DH10B NW20110620.A4

• E1: RP1@HB101 EM20110620.A1*
• E2: PSB3k3-P1010@DB3.1 EM20110620.A2
• E3: PSB3k5-P1010@DB3.1 EM20110620.A3
• E4: PSB3k5m(>-)-P1010 DB3.1 EM20110620.A4*

'*During plasmid prep, 1ml of Sol2 (instead of 300ul) added; user spun down samples, removed some of Sol2, before adding Sol3; asterisked samples already had some Sol3 added before spinning and removing (therefore some of the cells may have already lysed, the DNA was poured off)

Finalized construct design (see construct page)

June 24, 2011

Transferred cyanos to 10ml cultures in square flasks created Na2CO3 stock cryostocked norman’s plasmids (see 6/23/11) Silver’s S. elongatus isn’t doing so well Learned about gibson assembly, designed primers for constructs