miniPCR® P51™ Enzyme Lab: β-Gal Glow

Use an inquiry-based approach to investigate enzyme activity.

Investigate enzyme activity and the conditions that affect it. Under optimal conditions, enzymes catalyze, or speed up the rate of, chemical reactions. In this lab, a substrate that is not fluorescent under basal conditions will fluoresce under blue light when sugar molecules are removed through hydrolysis by the enzyme beta-galactosidase. Observing how brightly the solution fluoresces over time allows students to see the reaction progress and quantify the effect of different variables on enzyme activity. This lab includes four inquiry-based investigations. Students explore the effects of pH, temperature, enzyme and substrate concentration, and competitive inhibition on enzyme reaction rate. This lab provides enough reagents for eight lab groups to each pursue at least two of these investigations.

Required equipment (one per lab group):
- Micropipettes and tips.
- P51™ Molecular fluorescence viewer or other blue light transilluminator (460 to 480 nm, e.g. blueBox™ or blueGel™).
- Suggested for temperature investigations: Thermal cycler or heat block that can accommodate PCR tubes (ideal setup) or water bath and floaties, ice (workable).

Storage notes:
- Reagents require refrigerator storage.
- Reagents are stable for 6 months when stored in the refrigerator.

Note: Reagents are compatible with P51™ molecular fluorescence viewer or other blue light transilluminator (460 to 480 nm, e.g. blueBox™ or blueGel™).