Bacillus Studious

Conducting heat shock on E. coli to promote uptake of our ligated plasmid!

After conducting heat shock, we plated our bacteria along with a control bacteria which already had the Kanamycin resistant plasmid. By plating on Kanamycin, we can ensure that only transformed bacteria are grown. As you can see, quite a lot of our bacteria was effectively transformed since we had a LOT of growth on our ligation plate!

It’s super cool because you can see that there’s actual bacteria in the bottle!  I took this photo for my biology IA regarding aquarium biological supplements and their efficacy in establishing nitrogen cycles.  I gram stained a little sample of Tetra SafeStart to see the bacteria more clearly.

Close up photo of our gel!

Results!  The lane closest to the edge of the gel is a DNA ladder (or DNA standard).  Next to it is the solution we created in which we ligated the plasmid and kanamycin resistance gene.  And next to that is an unligated control solution containing the separated plasmids and kanamycin genes.

We’re working on transforming bacteria using plasmids this week!  First we ligated the kanamycin resistance gene to a plasmid, and now we’re testing to see if the ligation was carried out correctly using gel electrophoresis.  If we see DNA bands of 4,300 bp in our ligated solution, we’ll know that the ligation was carried out successfully since the plasmid is 3,000 bp and the kanamycin gene is 1,300 bp.  Sometimes the plasmids and genes get ligated together incorrectly so it’s important to check to make sure the solution contains viable plasmids for use in transformation.

Anyways, yesterday my Arabidopsis seedlings sprouted.  They are SO tiny.  I couldn’t help but squeal just a little at how cute they were.  Anyways, we’re essentially testing their salt tolerance right now, so I moved about 9 of the seedlings to a “high salt” plate in the tissue culture lab so I can compare the root growth of the plants in high salt vs low salt agar.  In total, about 80% of the seeds I planted germinated.  Sadly, the condensation in the plates ended up drowning a couple of them, though.  Nevertheless, I still have 9 good replicates for both the high salt and low salt groups.  I’m looking forward to seeing their growth tomorrow!

We also bleached our Arabidopsis thaliana seeds to prepare them for culture.  It’s amazing how 300 seeds can fit in such a tiny little portion of a microtube.  

Here’s a sample of some algae from my aquarium at 400x

We had our first A day this year today, which means I had Chem this morning.  We started working on a lab.  Pretty simple stuff really, just to help us learn about measurements and uncertainty and sig figs.  I can’t wait for AED tomorrow!  We’re learning about Arabidopsis thaliana.

In Bio we had “biology show and tell” where each of us brought in an object/picture/article that we thought was cool or that inspired us to become biologists.  I brought in a vial of river water from the river behind my house because I have always been super interested in marine biology and river ecosystems.  I had a great time watching people’s faces light up as they talked about what inspired them to be curious about science.  Good day overall.  

Seriously, getting new mildliners increases my urge to study by like 20000% percent

Using Christmas break to catch up on some Japanese :)