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:
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