User:University of Washington/25 August 2008

Conjugation BioBricks (Scott)
·PCR TrbA gene ·grow TrbC gene for minipreps and sequencing ·purify KorA gene for ligation tomorrow

LuxR from AraC and TetR(Faifan)
- got no growth from second time inoculating the transformation of ligated DNA(promoter + GFP/LuxR + pSB3K3)

- grew overnight I0462 from glycerol stock x 3 with Amp

- started cloning protocol from Ingrid (after digestion, will treat with CIP to remove phosphate from 5' end so the DNA won't re-ligate to itself.)
 * Restriction Digest: mix
 * incubated 37 degree Celcius for 2 hrs
 * 20 mins denature at 80 degree Celcius
 * stored digestion product in -20
 * ran gel on promo SP, S, P, and no enz: used 3 ul DNA, 2 ul dye
 * Bands of promo SP, S, P ran differently from no enz and were around at 3 kb as expected. However, there seemed to be light bands at higher than 10 kb ladder in SP lane which could indicate that not all plasmid were digested.

MG1655Z1(Faifan)
-none grew in four antibiotic plates ==> lost pCS26 with Kan, pACYC184 with Cam/Tet??, and some plasmid with Amp.. likely??

-grew overnight from the culture from Tsy culture#3 plate.

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