Part:BBa_K081014:Experience
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Part BBa_K572001 - RFP Generator with PcstA Carbon Stress Promoter
Applications of BBa_K081014
Experiment 1 - performed by Lorenzo Pasotti and Mattia Quattrocelli
This generator was contained into pSB1AK3. The constitutive promoter BBa_J23100 was assembled upstream of the generator using BioBrick Standard Assembly.
After ligation reaction we transformed 1 ul of ligation product into 50 ul of Invitrogen TOP10 and plated transformed bacteria on LB agar plates with Ampicilline.
15 hours after plate incubation at 37°C, we put the plate on the transluminator. Positive transformants glowed under UV rays (see figure).
We also picked up some colonies from the plate using a tip and infected 1 ml of LB + Amp. After 3 hours of incubation (37°C, 220 rpm) we watched 50 ul of bacteria with TRITC channel. Results are reported in the following figure.
We can see that our RFP generator actually works.
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