C I P E S
The success of the laboratory depends on the use of high-quality reagents, such as are available from Carolina Biological Supply Company. The recipes are organized in five parts: DNA Isolation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, and Cycle Sequencing. Reagents and stock solutions are listed alphabetically within each section. Follow the recipes with care and pay attention to cleanliness. Use a clean spatula for each ingredient or carefully pour each ingredient from its bottle.
If you are just beginning or have minimum time for preparation, consider using a ready-to-use kit that contains all reagents you will need for 25 student samples. The following kit was developed in cooperation with Carolina Biological Supply Company:
Human Alu Insertion Polymorphism AT
Catalog # 21-1230 DNA Extraction and Amplification Kit includes Chelex, proteinase K, Ready-to-Go Beads (in 0.5 ml tubes), primer/loading dye mix, and DNA markers.
Catalog # 21-1231 or 21-1232 DNA Extraction, Amplification, and Electrophoresis Kit includes all of the above, plus agarose, electrophoresis buffer, and either ethidium (1231) bromide stain or CarolinaBLU stain (1232).
I. DNA Isolation
10 ml. Store at room temperature (3 months).
Saline Solution, 0.9% Sodium Chloride (NaCl)
Makes 1000 ml. Store at room temperature (indefinitely).
Note: This solution will be used for a mouthwash, so insure that all glassware/plasticware is clean and free of chemical residue.
If solution is to be used immediately, it need not be sterile. However,
for long-term storage, we recommend sterilizing by autoclaving for 15
minutes at 121°C or passage through a .45 or .22 micron sterile filter.
A L U I N S E R T I O N P O L Y M O R P H I S M
DNA Learning Center, Cold Spring Harbor Laboratory
Noncommercial, educational use only.
Kits available from Carolina Biological Supply Company.