Team:Hawaii/Notebook/2008-10- 8

= Things we did today =

Triparental conjugation (cont.)

 * Grace


 * PCR'd BBpRL cultures to ensure that plasmid was retained
 * Plasmid was not retained (grown in SOB w/o selection = no reason to retain plasmid)
 * PCR'd BBpRL colonies #1 and 18 from plate to ensure plasmid present
 * Regrew RP1, BBpRL harboring cultures in 20ml SOB with selection (Kan50 and Sp100 respectively) and incubated at 37C overnight, with shaking (rpm = 250)

Verification of transformants

 * Grace


 * Colony PCR of transformants
 * Restreaked nrsg #6, prpgt #5, 7, 11

Construction of secretion device (cont.)

 * Grace


 * PCR'd and verified on EtBr stained 2% agarose gel:
 * J33207
 * nir+rbs
 * nir
 * rgt#1
 * Overnight RE digest:
 * HindIII/BamHI:
 * J33207
 * EcoRI/SpeI:
 * nir
 * nir+rbs
 * XbaI/PstI:
 * rgt#1

Prepared for sequencing

 * Grace


 * Colony PCR'd:
 * pgt #21
 * nrsg #1, 2, 15, 17 (from 10/2)
 * sgt #13 (from 10/2)
 * ExoSAP'd:
 * pgt #21
 * nrsg #1, 2, 15, 17 (from 10/2)
 * nrsg #6 (from 10/8)
 * sgt #13 (from 10/2)
 * prpgt #5, 7, 11

iGEM title/abstract

 * Grace & Margaret


 * Revised abstract
 * Sent in abstract, title, track selection to iGEM HQ

= Discussion =

= Quote of the Day = "History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson"