Final answer:
The procedure for agarose gel electrophoresis involves preparing a gel with agarose and buffer, staining it with EtBr, loading the samples, and then running the gel through an electric current to separate the substances for analysis.
Step-by-step explanation:
Procedure for Agarose Gel Electrophoresis Lab
The lab procedure for analyzing substances using agarose gel electrophoresis begins by weighing 1 gram of agarose into a corked Erlenmeyer flask. Next, 100 ml of 1x TAE buffer is added using a graduated cylinder. This mixture is then heated for one minute in a microwave, swirled, and briefly cooled under running water. Subsequently, 10 µl of ethidium bromide (EtBr) is added for DNA staining, and it is essential to wear gloves as EtBr is mutagenic. The mixture is then poured into a gel cassette, and a comb is inserted to form wells. Once the gel has set, usually after about 20 minutes, samples can be loaded into the wells along with a molecular weight ladder. The gel is run for 20 minutes at 100 volts. After the run, a gel documentation system is used to take a picture of the gel for analysis.
Other important details include the use of a tray placed into a chamber for running the gel, with the negative electrode near the samples and the positive electrode on the opposite side. Proteins can also be run on gels in certain cases, following specific preparation steps. Additionally, assays like the Ouchterlony Assay, which involves immunodiffusion in an agar gel matrix, show the versatility of agar in such lab processes.