In this lab students get hands-on experience with cloning ribosomal binding sites onto a plasmid, then observe its effects on transformed bacteria compared to a control by quantifying the expression of GFP in red and/or blue colonies. Next, they use a thermocycler and the Golden Gate Assembly (GGA) technique to clone 2 different ribosome binding sites into a plasmid with a reporter gene. Finally, they transform their GGA reactions into E. coli. For 8 groups of 2 to 4 students.

Advanced—For experienced high school and college classes; requires some technical skill.

Bring your biotechnology classroom instruction to the next level with a unique synthetic biology lab that focuses on the regulation of translation. Students get hands-on experience with molecular cloning when they add ribosomal binding sites (RBSs) to a plasmid and observe their effects on the expression of either a reporter gene that causes colonies to turn blue or one that causes them to glow red under UV or blue light. An additional reporter gene that causes green fluorescence protein expression is removed during the cloning process, providing students with another way to differentiate colonies containing original unaltered plasmid from those containing plasmids that have been changed during the closing process. This one-of-a-kind, colorful lab experience enables students to easily measure the effects of RBS sequences on bacterial protein production as they learn several of the core methods of recombinant DNA technology.

In this lab, students also use a method of recombinant DNA production called Golden Gate Assembly (GGA) to clone a weak and a strong RBS into either of 2 reporter gene expression units in a plasmid. They transform their GGA reactions into E. coli and observe the ability of each cloned RBS to support expression of the reporter gene. Students quantify reporter gene expression using photographs of their colonies on petri dishes with free, easy-to-use ImageJ software.

• Designed for 8 groups of 2 to 4 students
• Teacher guide includes paper model activity to help visualize molecular processes
• Comes with FREE 1-year access to digital resources that support 3-dimensional instruction for NGSS

Note: Order the kit with the perishable materials included or with a voucher to request perishables later at your convenience. Contact Carolina or return the voucher to request delivery of perishables.