Part:BBa_K3020000:Experience

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Typical DNA damage agents include nalidixic acid (NA) (Sigma), formaldehyde (formaldehyde, CH2O) (Sigma) and hydrogen peroxide(H2O2).The following non-DNA damage reagent was used as a negative control:kanamycin sulfate，KAN）（Amresco）,acetone,phenol. The reagent concentration and gradient were calculated and added to the 40 ml bacterial solution (LB) system according to the following induction volume.

DNA damage agent concentration gradient and induced addition

non-DNA damage agent concentration gradient and induced addition

The BL21 strain introduced with Part:BBa_K3020000 was cultured overnight in LB medium (AMP resistance) at 37 ° C, and inoculated into fresh LB liquid medium (AMP resistance) at 1:50 inoculation, and shake cultured at 37 ° C for 3 hours. The OD600 is 0.2-0.4. Add DNA damaging agents and non-injury agents with different concentration gradients. The concentration and dosage are shown in the above figure. Induction at 37 ° C 130 r / min for 2-3 h. Standard error was taken three times for each set of experiments.