User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/06/03
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June 3rd, 2010
1. TEAM MEETING:
My new goals are my old goals, I couldn’t extract the blue promoter from genomic DNA by PCR, now we’ll synthesize it, we’ll take only 50 bp from the original promoter of 86 bp, this 50 bp include the -35 an -10 boxes, one spacer between the two boxes and 2 inverted repeats. We also added 22 bp of preffix at the 5’ end, to prime a plasmid in which we’ll insert this promoter and a restriction site for SpeI at the 3’ end. Primer sequence is in the file “Minimum blue promoter.fasta” in the “Dropbox/iGEM_LCG_2010/WiFi Colli/Blue receptor” folder.
When I have the promoter, I’ll ligate it to a reporter (BBa_K145015) to test functionality. Before the ligation I have to insert a RBS into the reporter because it lacks it.