Inquiry-based experiments for large-scale introduction to PCR and restriction enzyme digests

Area of Science:

• Biochemistry
• Molecular Biology
• Genetics Laboratory Education

Background:

• Polymerase chain reaction (PCR) and restriction endonuclease digest are fundamental techniques in biochemistry and molecular biology.
• These methods are typically taught at advanced levels, demanding significant instructional time.
• There is a need for accessible, introductory laboratory experiments incorporating these techniques.

Purpose of the Study:

• To present two inquiry-based experiments for introductory laboratory courses.
• To integrate polymerase chain reaction (PCR) and restriction endonuclease digest into a single experimental design.
• To enable students to identify unknown DNA sequences using combined molecular techniques.

Main Methods:

• Developed two flexible, inquiry-based laboratory experiments for introductory courses.
• Combined PCR amplification with restriction endonuclease digestion.
• Students determined unknown DNA sequences based on PCR product size and digest patterns.

Main Results:

• Successfully implemented experiments in courses with 584 students, achieving an 85% success rate.
• Students showed improved understanding of experimental concepts and data interpretation.
• Enhanced general laboratory skills were observed in participating students.

Conclusions:

• Inquiry-based experiments integrating PCR and restriction digest are effective for introductory biochemistry and molecular biology labs.
• These methods can be adapted for various resources and large student numbers.
• The experiments successfully improved student learning outcomes and practical skills.