Frustration & Learning

In my second week in my inquiry learning workshop I have been frustrated, but I have also learned about frustration and how it interferes with learning.

During the second week we transformed some bacteria with the GFP (Green Fluorescent Protein).
Now I do teach Biology and I am familiar with the steps required to do transformation, but is something I have only done once before in another workshop, over 4 years ago.

Now the first part is easy. We are given plasmid DNA that has DNA from a Jellyfish. A Plasmid is a piece of DNA that is found in bacteria, but is not connected to the bacteria's DNA. The Plasmid can move in and out of the bacteria and this is one way that bacteria can pass around the genetic information that makes them antibiotic resistance. What molecular biologist have learned how to do is to take a plasmid from bacteria and splice in another gene. Now this is one way that they make insulin for diabetics. They take plasmids, splice in the gene making insulin, put it into a bacteria and the bacteria will make insulin. They grow billions of E-coli and these E-coli produce insulin.

The plasmid DNA that I received came from a jellyfish that naturally glows green.















Each plasmid only carries one gene and each had a different gene from the jellyfish. So some might of have the gene for making the protein that glows, one might of have the gene for making the slime on the outside of the jellyfish or any other part of the jellyfish. We first had to transform the bacteria, that is trick the bacteria into taking in the plasmid. Once the bacteria takes in the plasmid it will produce the protein that was on the plasmid.

So this is what I grew. The top 2 rows have growth, but too much. When you have this much solid growth its called a lawn. In the third row only the first plate had growth, but these were colonies, basically little dots of growth. It was the same for the 4th row as well.
















Me and my lab partner had success, we had one dish that grew bacteria that had taken in the plasmid with the Green Fluorescent Protein Gene! You can see the glowing green little colonies.
















What was frustrating? The Stoichiometry (calculations) needed to figure out the concentration of bacteria cells per sample on the plates with colonies. This is something I have never done. Fortunately for me one of the classes I teach I do show how to do conversions, which is similar to what I'm supposed to do. So I kinda know how to set it up. I was also lucky is that my lab partner new what to do. So she showed me what to do. Thats good and bad, I learn better when I puzzle it out myself.

The next step is verification. When you transform bacteria to produce a protein you need to verify that you have the correct protein inside the bacteria. So you go through a process where you take the plasmid out of the bacteria and place the plasmid DNA on an agarose gel to see the length of DNA that you are interested in. Basically you look to see if the plasmid in your transformed DNA is the one you wanted in the bacteria.

So what was frustrating about this? The steps to do this were complicated as well as the calculations to make all the solutions we needed to make to perform the experiment. Now in the workshop we first meet to discuss what we are going to do, then we do it. The moderator/teacher of our workshop was going over what we had to do. She was going over the instructions in a fast manner and assuming we knew all the reasonings why certain things need to be done. This was all so frustrating. I was trying to listen and write down instructions. That is something hard for me to do because when I write I am comprehending what I heard, but then I get behind in what is being said. SO at one point I got lost and looked up to see who I could ask for what I missed and I saw that everyone was lost. I teach, I know what the "lost-look" looks like. The moderator (a post-doc student) didn't know we were lost. She didn't know because some of the teachers were nodding their heads lie they knew what she was talking about, and when the moderator asked "any questions?" no one raised their hand.

Now the kind of person i am is that when I don't know something I NEED TO KNOW NOW! I raised my hand and asked "I'm lost, I think everyone else is lost, please raise your hand if you are on the same page as the moderator" and no one raised their hand. So we had to back peddle a bit and we figured it out.

This was a good experience because this is exactly what happens to students in a classroom. They get lost and they just nod there head and wait until the bell rings. Whats worst is that students will not raise their hand to say they are lost. They have learned by high school (through bad teacher experiences) that if your the one that raises their hand to critique something in class you, as the student, risk of being embarrassed or made fun of or even in trouble. So I learned to pay attention to how I give instruction and to gauge their facial/physically reaction. so I don't lose them.

So once we did that we went to the lab. I actually did partially figure out how to do some calculations for the solutions we needed. We went through all the chemical steps to separate the plasmid and reculture it. We actually did this part on more than 1 samples so we would have more than one plasmid sample to test. Then we cut up the plasmid for just the DNA for the gene we grew in the bacteria. Once we do that we use a micropippete tool to place the DNA samples in the agarose gel.

SO these are micropippetes, you use them to distribute microlitres of fluid. That is 1/1000 of a ml, very small amounts.


The idea is that you cut up the DNA into little pieces. Then you make a gel, a jello like substance, that is about 1/4-inch thick and rectangular in shape about 3 inches by 4 inches. The gel is made in a mold that creates 12 small wells, or holes, on one end of the gel. You then micropippete a small amount of one of the DNA samples into one of the wells in the agarose gel. Then it is placed in a Electrophoresis tank and an electrical current is applied to the gel. DNA is polar, which means it has a positive end and a negative end. So when the electrical current is applied to the gel the DNA moves within the gel. This makes the DNA bands that TV uses in all there CSI shows. This is the Electrophoresis tank.


This is what we got after we took the gel out and put it in the UV ray to see the bands. IT WAS WAY COOL!

This is all we could do this week. next week will run a PCR, a way to reproduce the gene of interest in great amounts.

We ended the day having a discussion on inquiry learning and how we could incorporate the inquiry process in our classes. All teachers in my group realize that what we are doing would not be entirely possible in a basic biology class. Most of us, including me, do not have the tools or equipment for most of what we are doing. But we do take away our experiences with these procedures and an understanding how important it is to allow students to dig and figure out things on there own. This way they are engaged and when they are engaged they are learning.