Hi everyone! So, unfortunately, we got some disappointing news today. As it turned out, the gel that we thought looked so good last night didn’t actually look as good as we thought. Although there were bands present in both sample lanes, they weren’t in the right place (so they weren’t the right length), which means the ligation didn’t go well. Since we only have two weeks left, at this point we have to scrap the culture-independent portion of our experiment, which is disappointing since we put so much effort into it, but that’s science! Many of our culture-dependent transformations seem to be growing well, so later tonight we’ll go in and do pre-plasmid purification preparation (try saying that five times fast) on them, so that we can do a Qiagen miniprep and run a restriction enzyme digest on them tomorrow afternoon.
Since we’re done with the culture-independent project and our culture-dependent plates are still incubating, today we focused on staining bamboo stem and leaf sections. The sections turned out really well, so that was exciting! We used three different stains today: Toluidine Blue O (TBO) which is a metachromatic stain, meaning that you can apply this single stain to one sample and it’ll stain multiple features of the sample different colors. It stains pectins (sugars) purple, lignins (found in wood) blue green and cuticle dark blue. We also stained with Alcian Blue, which stains cellulose (in cell walls) bright blue. The final stain we used today was Safranin, which stains lipids and waxes reddish orange, and lignin dark red. The final stain we’ll use later next week is Phloroglucinol, which stains xylem bright red, but you need to use it with hydrochloric acid, so we won’t use it until later. We got to view the sections we did stain with a microscope in the basement that allows us to take high-quality photos, which I’ll include in this post—they turned out really well, gave us a lot of information about the anatomy of a bamboo plant, and will look good on our poster!