Watch PCR product glow with the naked eye — no need for gels!
Students will perform a modified version of a quantitative polymerase chain reaction (qPCR). qPCR monitors the amount of PCR product produced in real-time using fluorescent dyes or probes. This lab modifies the reaction so that students can observe the change in fluorescence first hand, without the need for a qPCR machine and using low cost equipment.
This activity serves as an excellent illustrative learning tool for how fluorescence and PCR can be used to quantify nucleic acids while also providing an introduction to the process of PCR in general.
- Techniques utilized: Micropipetting, PCR, quantitative PCR, gel electrophoresis (optional)
- Time required: can be completed in one 120 min. block, or extended into three class periods
Level: Intermediate to advanced.
The P51™ qPCR Lab kit contains reagents for 8 lab groups of up to 4 students each (32 students):
- 2X qGRN Master Mix
- qPCR Lab Primer Mix
- DNA Sample
- Nuclease-Free Water
- 100 bp DNA Ladder, Load Ready™
- 8-tube PCR strips
- 6X DNA Loading Dye
- miniPCR™ or other conventional thermal cycler
- P51™ Molecular Viewer or blue light transilluminator (460-480 nm, e.g. blueBox or blueGel) — one per lab group
- These reagents require frozen storage.
- Shelf-life: 12 months when stored at -20ºC.
Free miniPCR Learning Lab™ downloads:
Contact firstname.lastname@example.org for answers key.