Regulatory

Part:BBa_K5097000:Experience

Designed by: Kelly Acen, Marie Box, Alexia Darby, Jena Dookie, Blake Dieckman, Runya Chaora, Chris Dorce, Adrianna Dugan, Elber Lopez-Hernandez, kayla VanPelt, Makenna Ventuleth   Group: iGEM24_Oneonta   (2024-09-30)


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Applications of BBa_K5097000

Experience

We used this riboswitch sequence to design a reporter construct (Bba_K5097003) to measure the pH responsive expression of BFP under the control of PRE. After cloning, we screened colonies by colony PCR, identifying many positive clones.

alx-pre-bfp-psb1c3.jpg
Figure 1: Colony PCR confirms successful cloning of PRE-BFP into pSB1C3. Bacterial clones were then subjected to colony PCR and analyzed by agarose gel electrophoresis. The ~1200 bp band indicates the positive clones.

Positive clones were then mini prepped, and a sample of DNA was sent for Sanger sequencing to confirm the riboswitch sequence prior to whole plasmid sequencing, Unfortunately, we were unable to find a clone whose DNA sequencing result indicated a complete and correct PRE riboswitch sequence.


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