Generator

Part:BBa_K081014:Experience

Designed by: Lorenzo Pasotti, Paolo Magni   Group: iGEM08_UNIPV-Pavia   (2008-10-19)
Revision as of 03:06, 6 October 2011 by Domit (Talk | contribs)

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

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.

Pv RED.jpg
Pv REDmicro.jpg

RFP Generator with PcstA Carbon Stress Promoter

User Reviews

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