Cloning and Sequencing Explorer Series

The complete Cloning and Sequencing Explorer series is a 6–8 week series of laboratory activities in which students clone and analyze a portion of the GAPC gene. Activities range from DNA extraction to computer-based sequence analysis.

The first step in a gene cloning lab, the Nucleic Acid Extraction module allows students to extract all nucleic acids from plant cells using tissue grinding and lysis, followed by purification using column chromatography.

An advanced PCR lab that utilizes nested PCR, the GAPDH PCR module allows students to amplify a portion of the GAPC gene, a housekeeping gene in the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) family, from a plant of their choice.

These DNA electrophoresis reagent packs are available in small, medium, and large sizes. Students can pour their own gels to visualize the products of PCR reactions, ligations, and restriction digests or perform dye electrophoresis.

The PCR Kleen Spin Purification module uses size exclusion chromatography to remove unincorporated primers, nucleotides, salts, and enzymes from PCR products.

This module allows students to directly ligate purified PCR products into the pJET1.2 plasmid vector and immediately transform freshly prepared competent bacteria with the ligation reaction.

The Microbial Culturing module contains the materials necessary to teach basic sterile technique, culture and transform ampicillin-resistant E. coli, and grow cultures for plasmid minipreps.

In this module, students can rapidly purify up to 20 µg of plasmid DNA from transformed bacteria using a three-step alkaline lysis procedure followed by mini column chromatography.

The sequencing and bioinformatics module is used to analyze a portion of the GAPC gene isolated from a plant. Students are guided through a sequence analysis that allows them to make a real contribution to science!