P51™ qPCR Lab: Principles of Quantitative PCR
Perform quantitative polymerase chain reaction (qPCR) in a directly
observable format and understand molecular diagnostics. qPCR monitors
the a#scent dyes
or probes, and it is the gold standard for the detection of viral
infections including COVID-19. In this lab, students use low-cost
equipment to observe the change in fluorescence first hand, without the
need for a qPCR machine. Students directly follow the increases in
fluorescence that accompany PCR cycles, understanding how exponential
DNA amplification works. They then use their measurements to quantify
nucleic acids. This lab grounds students in the principles and practice
of qPCR, the gold standard technique in the molecular diagnosis of
infections including SARS-CoV-2. An optional extension allows students
to compare results of real-time qPCR and end-point PCR using gel
electrophoresis.
- Techniques: Micropipetting, fluorescence detection, quantitative PCR.
- Topics: Gene expression, fluorescence, biotechnology
- Time required: 120 minutes; can be split over 2 or 3 class periods
- Level: Advanced high school through college
What is included
The P51™ qPCR Lab kit contains reagents for 8 lab groups:
- 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
*Note: Reagents are compatible with P51™ Molecular Fluorescence Viewer or other blue light transilluminator (460-480 nm, e.g. blueBox™ or blueGel™).
Required equipment (one per lab group):
Optional extension requirements (end-point gel electrophoresis):
Storage notes:
- Reagents require freezer storage
- Reagents are stable for 12 months when stored in the freezer
If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200