{ "date" : "2013-09-02", "author" : "emil", "title" : "50 shades of red(FP)", "content" : " I have a new aim, to determine which is the best chassy between 5α ,10β, TOP10, TB1, JM109, BL21, I'll transform all that one with the same construct pSB1C3 with pLac and mRFP as reporter gene, I'll do the inocula and when the O.D. will reach 0.7 I'll induce with different concentration of IPTG, this could be useful for the modeling page(0.025 mM; 0,05 mM; 0.075 mM; 0.25 mM; 0.5 mM; 0.75 mM; 1 mM; 1.25 mM; 2.5 mM).
After that I'll take 1ml sample at every hour for 5 hours.
Finally I'll centrifuge every sample in order to replace the LB with PBS then I'll sonicate every sample for 10 seconds and after a new sonication I'll transfer the surnatant in glass cuvettes for the analisys with the fluorometer! The mRFP adsorbs at 584 nm and emitts at 607 nm. This is a real long work and this is my LAST POST!", "tags" : "E.coli strains" }