pGLO Lab







Two new traits in the bacteria consist of glowing green colonies and ampicillan.

The amount of bacteria that would be in -pGLO LB would be none. The amount for bacteria that would appear in -pGLO LB/Amp is none. The last to are different though as +pGLO LB/Amp would produce around 448 colonies and +pGLO LB/Amp/ara would produce 640 colonies.

The role that the arabinose played was that it attaches to the promoter and then triggers the glowing.

GFP is used in mice. They tried it in mice to test if they can get the florescent look they get in jellyfish an found that the mice glowed everywhere but its hair. They also tried it in plants to test if they were freezer resistant, if the plant glows you know that the plant is freezer resistant. They have even made fish with GFP in it. These fish glow and are actually sold to people. The fact that the fish glows will appeal more to a certain demographic, kids.

An example of another application of genetic engineering would be to figure out if people are related. So for example you can take their genetics and compare them to each other.

Thinking Like a Biotechnician 

Today we conducted a lab that showed the imitation and the process of inserting plasmid into a bacteria plasmid. Our plasmid had a resistance to ampicllin. As part of this lab we were told to find a restriction enzyme then cut a segment out of the DNA and cut the plasmid open. The enzyme used was closest to an insulin gene. We used an enzyme that was closest to the insulin gene. My group then combined the DNA and plasmid with Ligase. We used tape to represent the Ligase. If we had used kanamycin or tetracycline as our resistance it would have kill our bacteria, so in order to test if bacteria took our plasmid, we used ampicllin because our plasmid has a natural resistance to it and if the bacteria survived then the insertion would be successful. The enzyme restriction that we used to cut the DNA segment was called Hin III. We used Hin III because it cut the the human gene in two places and cut the plasmid in only one 1 place. The reason for this criteria is because if the restriction enzyme cut the bacteria in more than one place, then the lyase would not know where to attach the insulin gene to. This is important in every day life, because it is used to create many known antibiotics and show, so that we can test and use these products. This technology can also be used today to create longer lasting foods or help kill viruses.

SMART Goals

In this new semester of Biology I would like to make a SMART goal that can better improve my performance in the class. I am motivated to come up with better study skills and routines. I will work towards practicing how to study and then put the skills into action. My action plan will be to practice study skills for 10-15 mins every time I complete a vodcast. I will study the content of the vodcast I just completed using the given study tips by Mr. Orre last semester.

I would also to present another SMART goal which is not biology related. This goal has to do with reaching out to friends. I find that sometimes my friends feel frustrated with me because I do not reach out to them outside of school. I will when time isn't of the essence, call or text a friend and ask them if they would like to hang outside of school.

Protein Synthesis Lab

          This process is call protein synthesis. A copy is made of one side of the DNA (called the mRNA) where the gene is located. This copy is transferred to the cytoplasm. Then mRNA then leaves the DNA and travels to the ribosome. The segment is fed through the ribosome. The mRNA bonds with a ribosome, which will make a protein. The ribosome reads the first three bases of the whole sequence where the ribosome translates the amino acid (it will repeat this action through). The mRNA bonds with a ribosome, which will make a protein.

          When a mutation occurs it can either be a mutation that makes a small difference or a big difference. A small problem occurs when a mutation effect the amino acid sequence but in total the mutation does not make the base a different one. A big difference is when a base is added or pulled out and the amino acid changes. There are two main topic kinds of mutation. Substitution which is when a nucleotide is substituted for another. The other, frameshift mutation - a frameshift mutation breaks into two separate parts. Those two parts are called insertion (when an extra base pair is put into the code) and deletion (when a base pair is left out of the code). If a T was inserted into the code at the end versus the beginning it would do less damage to the sequence. This is because the mistake acts like a chain effect. 

           In my own sequence on for the lab I chose to add in a frameshift mutation because I though this would make the biggest effect. I inserted an extra base in the very beginning, specifically the fourth base. This then effected the rest of sequence. It matters where the mutation occurs because it can either effect the whole rest of the sequence or only some of it.  

          Amino acids can really effect you life. For example there is one mutation that happened and its called Apert syndrome. Apert syndrome s a genetic disorder characterized by the premature fusion of certain skull bones and sometimes even fuses fingers and toes together. Almost all cases of Apert syndrome result from new mutations in the gene, and occur in people with no history of the disorder in their family. Although if one has Apert syndrome they may pass this on to there newborns.

Unit 5 Reflection 

This unit was about coping DNA, and translating it. It is called walking the dogma. We talked about the difference between DNA and RNA. RNA is single stranded and has ribose - not deoxyribose, and RNA contains unracil instead of thymine. Mutions was also brought up and we look at the different kinds of mutations, there were two main of mutations. One of which is called substitution, this is when one nucleotide is substituted for another. The second kind is called a frameshift mutation and in a frameshift mutation there are two kinds that branch off. Insertion (when an extra base pair is added to the code) and deletion (is when a base pair is left out of the code). I think my strength was based around mutations because I liked learning about how much a mutation could really effect you. Usually when I enjoy something I excel at it. We brushed over the topic as to why we should care about mutations. We said that it changes DNA - more specifically it changes proteins which are essential to life. Some diseases are caused by a change of a single base! Mutations in DNA can lead to cancer and they can create new genetic variation giving population new traits for natural selection to act on. We looked at gene expression and regulation. Gene expression is the process of a gene begin used to produce a gene project or phenotype. A gene regulation is a mechanism used by cells to increase or decrease the expression of a gene. This is where I ran into trouble as I feel it go a

lot harder trailing off into promoter, operons, and operaters. I learned that I am usually better at something that I enjoy and can ask a lot of questions, after I watch the vodcast I usually find myself pondering over questions i've asked myself. I think that I have grown as I biology student but I have also learned a lot more about myself and my learning style.

Human DNA Extraction Lab

        In this lab we asked the question: "How can DNA be separated from cheek cells in order to study it?" In this lab we found that you can separate DNA from your cheek in order to study it. I found that in this experiment you can see DNA when you follow three basic steps: homogenization, lysis, and preciptiation. You can see the DNA when you add detergent, salt, enzyme and cold alcohol to create a gatorade solution. I observed that my DNA floated to the top once I had added the cold alcohol. I looked as though bubbles carried it to the top. This observation helped me strengthen my thinking and thoughts that came out of notes and vodcasts I watched. This data supports my claim because DNA extraction is the process of homogenizing (to combine two unlike things), lysis (a process of disintegration or dissolution (as of cells)), prectiptaion. When the protease (the enzyme) was added to the gatorade solution outcame a faint reaction and the DNA was in the making, but when the layer of alcohol was added the DNA was made visible in the test tube.
         While my hypothesis was supported by my data, there could have been errors due to me not scraping my cheek enough when I was swishing plain gatorate in my mouth. I know this was an issue because group member who had kept the gatorade in there mouth longer and scraped there cheeks more effectively had more DNA. In order to eliminate this error next time I would scrap my cheeks more to break off the cells. A second error that could have affected my experiment was adding too much gatorade to the solution and not enough alcohol. Adding to much gatorade also makes the DNA less concentrated and could have affected the over all process. To prevent this from happening again I would use less gatorade to balance out the amount of alcohol that was added. Due to these errors, in future experiments I would follow my recommendations in order to avoid anything affect your experiment.
         This lab was done to demonstrate that DNA could be separated from cheek cells in order to study and observe it. From this lab I learned and what an enzyme help create a reaction an eventually some DNA, this helps me understand how a enzyme works on a larger scale. Based on my experience from this lab, I could apply this knowledge to another situation if I were to maybe look at DNA on more of a molecular level, and needed to re-create someones DNA in that kind of form.

Egg Lab Conclusion 

             In this lab we asked the question: Can macromolecules be identified in an egg cell? 

Our claim/answer to the question was that, yes they can but they may not be identified in all macromolecules. Also, in the egg membrane the macromolecules that were present were polysacchrides, lipids, and proteins. In the egg white the macromolecules that were present were proteins, polysacchrides, and monosacchrides. In the egg yolk the macromolecules that were present were the lipids, proteins, monosacchrides, and polysacchrides. Each test had a chemical added to it, for us to see if the macromolecules were present. There were four different indicators; Benedict Solution, Iodine, Sudan III/IV and Sodium Hydroxide with Copper Sulfate. Evidence that supports our claim is as follows: In each test we would know if the macromolecules were present because the test would change to a darker color. We were asked to rate how present the macromolecules were out of 10. We found out that the membrane has polysaccharides because the solution turned black and it's found on the surface of the cell membrane, I rated this a 5/10. The membrane also has lipids, the solution turned orange, and the membrane is made of phospholipids, which is a lipid, so this makes sense. I rated this a 2/10. Lastly, the membrane showed the macromolecule; proteins, and in effect the solution turned dark purple. I rated this a 6/10. All in all, the egg membrane showed the macromolecules, polysaccharides, lipids, and proteins. We also found out that the egg white has polysaccharides(3/10), monosaccharides(7/10) and lipids(4/10) for energy growth and development. The egg white contains proteins for two purposes, first one is for growth and development, but also as enzymes, to protect it from bacterias. We know that the macromolecules were present because when we did the different tests the colors of the indicators because darker, just as the egg membrane test. Lastly, the egg yolk contains all four macromolecules, it contains monosaccharides(3/10), water(8/10), polysaccharides(4/10) and lipids(7/10) as energy, but also to make up the cell's internal membrane, which is covering the yolk. Again, the indicators colors turned a darker color to show that the macromolecules. The Benedict solution turned a darker navy blue. The Iodine turned a dark brown/purple. The Sudan III/IV turned a dark red/orange. Lastly, the the copper sulfate turned a dark royal blue. This data supports our claim because it one figure out which macromolecules are present in the different parts of the egg.

          Some possible errors that could have happened are the amount of drops of each solution where dropped into the different tests. My group dropped in 1-2 drops. This could have effected the outcome depending on whether my group members dropped in 1 or 2 drops this could have effected the outcome by a little. Another possible error is the amount of ml (the parts of the egg we tested. which were in the test tubes, they may have not been the right amount and it may have affected the color. To improve this lab, I would first have a perfectly controlled and tested experiment where only one person does everything so you know how many drops to do, or just tell your group mates to use the same amount of drops. To solve the second problem,  I would have used another tool to get the 1 ml of water, white, yolk and membrane, to be more precise.

           The purpose of the lab was to demonstrate that different macromolecules are present in the different part of the eggs. From this lab, I learned that certain macromolecules were only present in certain parts of the cell and that they had different uses depending on where they were, which helps me understand the concept of macromolecules and their functions. Based on my experience from this lab,  I may be able to do more experiments with eggs, and solutions such as the indicators that we used in the lab. Taking other food items and adding indicators to the different parts and seeing whether or not it has any macromolecules.