Safety
In addressing the safety issues, teachers explained the lab safety rules to us and supervised the whole process of the experiment. The project was performed mainly in open bench and fume hood. Lab coats, gloves, safety googles, and ethanol are used for protection.
Here are several safety measures concerning our experimental procedures:
1. Polyacrylamide gel electrophoresis (PAGE)
PAGE is used to observe the binding between each strands and the target RNA oligos. Since the acrylamide used is neurotoxin, we wear gloves to handle the solution and the gel. Also, the TEMED used in making the gel will cause digestive and respiratory tract burns, it is harmful If it is inhaled. Thus, the gel is made inside fume cupboard.
Moreover, for the DNA stains we used, since ethidium bromide is carcinogenic, it is dangerous for us to handle it, so a safer alternative, SYBR safe is used in our experiment to post-stain the polyacrylamide gel.
2. Peroxidase activity assay
When testing for the peroxidase activity, hydrogen peroxide which is a strong oxidizer is handled with care. Gloves and safety googles are put on when diluting the solution.
3. Cloning
We chose E. coli (NEB® Stable Competent E. coli) as our chassis as it belongs to risk group 1 which will not cause disease in healthy adult human. Thus, it is rather safe for secondary school students to handle. Gloves, safety googles, Bunsen burner are involved in the cloning experiments. The tools contaminated by E. coli are treated, e.g. autoclaved, before disposal.