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Team:Bielefeld-CeBiTec/Results/Biosafety/Long-termStability
From 2014.igem.org
Biosafety - Antibiotic-free Selection
Long-term stability of the antiobiotic-free selection
To verify if the plasmid stability of the antibiotic-free selection system is comparable to that of an appropiate antibiotic, the plasmid BBa_K1465401 was cultivated for 40 h in LB media, containing normal LB, LB supplemented with 5 mM D-alanine and LB containing 30 mg/L Chloramphenicol. When stationary phase of the cultivation was reached the cultivation was diluted to OD600 of 0.0001 and then diltued again 1:1000. From this dilution volumes of 10 µl, 20 µl and 50 µl were plated out to estimate the ratio between red colonies (harboring the plasmid) and white colonies (loss of the plasmid).
The cultivation with 5 mM D-alanine should probably led to the loss of the plasmid, because it is not neccessary for bacterial grwoth when D-alanine is supplemented. And as shown in Figure 10 below the
Prozedere erläutern
Resultat über 40 h ist antibiotika-frei gleiche Plasmidstabilität gegeben wie mit Cm, aber abgebrochen wegen Zeit, aber eigentlich auch unnötig, da solange nicht damit kultiviert wird...
