cDNA Sequencing By Restriction Project

Teacher: J. Kirk Brown
Subject: Biotechnology
Grade Level(s): 9 10 11 12
Target Audience: Biology II, AP Biology and Genetics
Materials Needed: microcentrifuge, LB Amp Agar, broth, electrophoresis equipment, restriction enzymes
Class Time: 3 periods
Brief Summary: This project is an actual part of the Human Genome project. Students are actively engaged in trying to confirm the sequence of a given cDNA clone using the WWW tools and traditional restriction analysis. The students grow the clone, prep the plasmid, cut the plasmid with restriction enzymes and compare their results to what was predicted to cut via webcutter on the world wide web.
Student Objective(s): Students will use the WWW as a tool in biotech/genome research. Students will conduct real research on an actual cDNA clone provided by Lawrence Livermore National Laboratory. Students will learn miniprepping, electrophoresis, WWW use and clone growth.
Integration (tying it all together): This activity integates computer technology with wet lab techniques. Students see the computer as a vital link in the biotech lab.
Description of Activities: The students are provided a stab culture of clone 26962 (human cDNA). They use webcutter on the WWW to predict how many times a chosen restriction enzyme will cut the tentative sequence as provided by D. Greg Lennon at Lawrence Livermore National Lab. Then they perform the experiment and compare actual vs. expected DNA fragments. They then share the information with others. Each school should try different enzymes.
Further Information and References: High school cDNA sequencing by restriction project. http://www.tracy.k12.ca.us/cdna/cDNA_project.html

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