IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/08/10

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August 10th, 2009

Run PCR to confirm genomic integration at specific promoter regions.

Dilute Primers for this

Restreak E.coli onto LB/AMP for DNA replication

Primer Dilutions Stock Concentrations Wanted Concentrations Directions
YHL012W-Check 100mM 5 mM take 5 uL of primer and dilute it into 100 uL of TE buffer
YGR287C-Check 100mM 5 mM take 5 uL of primer and dilute it into 100 uL of TE buffer
YDL243C-Check 100mM 5 mM take 5 uL of primer and dilute it into 100 uL of TE buffer
YGL205W-Check 100mM 5 mM take 5 uL of primer and dilute it into 100 uL of TE buffer
YJL219W-Check 100mM 5 mM take 5 uL of primer and dilute it into 100 uL of TE buffer

PCR Design 25 ul RXNs, on bead, one blot of cells

PCR Tube #s

1 2 3 4 5 6 7 8
Strain Number and Name #1, YJL219 2#4 YGR287 2#6 YGR287 3#4 YDL243 3#6 YDL243 4#1 YHL012 4#2YHL012

Run PCR using superfolder E-COli as template. Hopefully get superfolder fragments which can be ligated onto the longtin plasmid. If not, I innoculated a couple of Falcon tubes of LB/AMP with superfolder E.Coli