Team:Groningen/Notebook/Wetwork 6August2012
From 2012.igem.org
Emeraldo
Another attempt on inserting the terminator with biobrick suffix and prefix inside pSac-Cm MCS. Terminator+prefix and suffix PCR, cut both original pSac-Cm and PCR product with EcoRI and HindIII, ligation reaction, and then E.coli DH5alpha transformation.
Tom
PCR alsT (150, 250, 300, 500) + GFP construct with taq-polymerase.
Restriction analysis for alsT (150, 250, 300, 500) + GFP with ecoRI.
Renske
Counted TAMC plates + analyzed results:
Arjan
GC-MS
Did liquid injection of rotten meat volatiles using methanol as organic solvent on the HP-5 column only. This methode yielded a spectra with multiple peaks and database reference match.
reference
Renske
Primer design for positivie feedback loop (with Nisa)
I PlacI
a) Prefix + PLacI_Forward
b) homology with pRe_F + PLacI_Rv
c) homology with pLuxR + PLuxR_Rv
II P_Activator
a) homology pLac + PRE_F
b) homology pLac _ pLuxR
c) homology cII with pRE_R
d) homology LuxI + pLUxR_R
III Activator
a) homology with pRE + CII_F
b) homolgy with pLuxI + LuxI_F/ GFP + CII_R/GFP+LuxI_R
IV GFP
a) CII + GFP_F
b) LuxI + GFP_R
c) suffix _ GFP_R
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