Getting more Phun Part 2…
My enriched isolation i mentioned in “Not so Phun Part 1”, yeah. That didn’t work out. But, thankfully, I think I found what was going on: The procedure indicated that we had to open the enriched isolation tube every 24 hours for air to get inside. however, we did not, so anaerobic bacteria started to grow, and most likely denatured my phages. The group of people doing the enriched isolation with me also did not yield any phages. My lab partner, Jake, found phages, BUT it was because he had used a flask with an open top, so that air could get in. I did not use a flask at first, because there weren’t any left.
Proof of Adoption:
After determining what went wrong, and why I could not yield a phage, I had the choice of going through the enriched isolation process again. However, I instead chose to “adopt” a phage from Jake, in hopes of saving time. I labeled 4 plaques I identified from his old plate (he had not streaked these plaques yet), and streaked them on agar plates. Hopefully, these streakings will confirm that these plaques are indeed phages when I check back in two days.
Well, two days after, my plates indeed had plaques… but it was horribly infected. Luckily, there were some noticeable colonies that did not seem to have touched the contaminants, so i streaked from those, and hoped for the best. I also streaked from an extra not-so-contaminated plaque and filtered it with a 0.22 micrometer filter. After plating all those new streaks, I came back five days later to completely contamination-free plaques. The method worked, and I can finally go onto streaking for my generation 2 phages.
No more contamination!!: