Team:Hawaii/Notebook/2008-09-25

= Things we did today =

Verification of Ligation

 * Margaret


 * The plac/rbs+rep and oriTr(BB) transformations resulted in 0 colonies with the correct insert, so the remainder of the ligation reaction from 9/16 was digested with XbaI and run on a gel.

Sequencing & Colony PCR

 * Margaret


 * PCR
 * oriV colonies (3,4,6,8,9,10,11)
 * aadA(BB) colonies (4,5,7,8)
 * aadA(pRL) colonies (1,2)
 * oriT + J33207 (1,2,4)
 * oriT + J33207(de-phosphorylated) colony 1
 * 5 additional colonies of oriTr transformation were PCR'ed

Construction of secretion device and redo of BB-pRL1383a (cont.)

 * Grace


 * Ran RE digests from last night on gel
 * nrsg8 and nrg4 did not cut. Implies insert wrong?
 * Extracted bands from gel
 * Ligated:
 * 3A using pSB1A3:
 * nrg7 + tt
 * rgt1 + nir
 * rgt1 + plac
 * rgt2 + nir
 * rgt2 + plac
 * pSB1A3 (control)
 * pRL1383a + J33207
 * pRL1383a to self (control)
 * Transformed into DH5&alpha; using 5 &mu;l ligation reaction; also transformed J33207 and pRL1383a plasmids as control)

= Discussion =

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