Bacterial with plasmid insert was selected by Ampicilin in glycerol stock and keep it in -80 deep freezer. BUT I streak bacterial from glycerol stock outside lamina flow hood. Do you think there is a chance my glycerol stock will be contaminated? Do I need to recheck it again e.g. check plasmid size, insert size?
Chuck A Arize
Check for Contamination: Before using the stock, streak a small sample on a selective agar plate. If you see growth that differs from your expected colony morphology, contamination may have occurred. Verify Plasmid Presence: It’s a good practice to verify the plasmid size and insert size using techniques such as PCR or restriction digestion followed by gel electrophoresis. This will confirm whether the plasmid in your glycerol stock is intact and contains the correct insert. Maintain Sterility: For future handling, always use aseptic techniques when working with glycerol stocks to minimize contamination risks. If you need to work outside of a laminar flow hood, be sure to thoroughly disinfect surfaces and use sterile equipment.
Yes, there is a chance that your glycerol stock could be contaminated, especially since you streaked bacteria from the stock outside of a laminar flow hood. Working outside of a sterile environment increases the risk of introducing contaminants, such as bacteria or fungi, into your samples.
To ensure the integrity of your glycerol stock, it’s advisable to recheck it. Here are a few steps you can take:
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