PCR Cycle Number Analysis MiniLab

Model exponential growth using polymerase chain reaction (PCR) and visualize with electrophoresis.

Description
How do you take a very small DNA sample and still be able to diagnose a patient with a possible disease, study specific genes at their base-pair level or solve a crime? You make copies of it! The discovery of Polymerase Chain Reaction, PCR, took an in vivo process of DNA replication and made it an in vitro system, transforming our ability to study DNA.

In this hands-on lab you will run PCR reactions for various number of cycles to see at what point you have generated enough copies to detect the PCR products by gel electrophoresis. This activity will illustrate the natural phenomenon of exponential growth, allow your students to model the copy number of DNA mathematically during PCR, and see the results of their predictions.

Materials Included in Each MiniLab
Each MiniLab contains enough materials for 10 workstations, 2 – 3 students per workstation. Materials include:

• Ten 1% agarose GreenGel™ Cups
• One bottle of 100 mL Tris-Borate-EDTA (TBE) buffer concentrate
• FastTaq PCR MasterMix (2X)
• Forward and reverse primer set
• Lambda phage genomic DNA sample
• MiniOne® DNA marker
• MiniOne® 5X sample loading dye
• One bag of 0.2 mL thin-wall PCR tubes
• One bag of 0.65 mL microcentrifuge tubes
• Teacher guide