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From 2012.igem.org

Arjan/Renske

Micro-Array cell harvesting

O/N culture diluted to OD = 0.326 (9:20h). Grown until OD = 0.775 (11:05).
3.6 mL brought to flask with magnet stirrer and 45 mL LB, rest put back in 37 degrees shaker for next experiment. OD = 0.095. 11:11 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over fresh meat* (kept on ice).
At 13:05 cells harvested by centrifuging (1 min, 10000 rpm, 4 degrees C) and put in -80.
12:20: remaining culture (OD = 1,78) diluted to OD = 0.311. 2 hours, 37 degrees (OD = 1,2). Brought to to flask with magnet stirrer and 45 mL LB, OD = 0.118. 13:20 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over rotten meat* (kept on ice).

*minced meat, 30% beef, 70% pork, bougth at Jumbo grocery store. Rotten meat was kept for 14 days in fridge, fresh meat was bought today.

Emeraldo/Nisa

1.) Plasmid isolation of BBa_I742123; concentration: 280ng/ul
2.) Linearization of isolated plasmid with SpeI. Result: correct length
3.) B. subtilis transformation with: BBa_K090403 (amyE integration backbone), BBa_I742123 (multi host vector plasmid), BBa_K274002 (violacein pigment). The yellow pigment and red pigment are not used here, because of the amp resistance gene: B. subtilis can't receive amp resistant gene. (the pigment genes must be inserted to another backbonoe with different antibiotic resistance).

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