pGLO

pGLO Observations , Data Recording & Analysis

Plate Number of Colonies Color of colonies under room light Color of colonies under   UV light - pGLO LB carpet white white - pGLO LB/amp 0 white white + pGLO LB/amp 150~ white white + pGLO LB/amp/ara 70~SO white green glowing(only 30~ colonies glow)

	In our test, the bacteria gained the traits of ampicillin resistance, and the pGLO gene, which makes them glow green when exposed to arabinose sugar.
	We spread 100uL of bacteria on the petri dish, and in that 100uL estimate that there are roughly 1500 bacterium in 100uL because there are 150 colonies on the the one with ampicillin ,and the gene. There are at least to times as many bacterium on the plate with only lori broth, so the must be atlast 1500 bacterium in the 100uL.
	The arabinose sugar allows the GFP gene to be repressed. When it is present, it attaches to the repressor for the GFP gene, and prevents it from attaching to the GFP operator, which allows the GFP gene to be expressed. The GFP gene can be used to tell if an organism has also taken a gene the will not be expressed. You can tell if it absorbed insulin gene, if it glows green. It can also let you follow the movement of substances in an animal, and track the progress of cancer through a brain. One other example of genetic engineering is our turkeys. The have been bred over the years to become as plump as possible.

Candy electrophoresis Lab

In our experiment, all of the dyes moved in the same direction, lined up with a reference dye, and was both the same size, and color as a dye.



Yellow 6, Red 40, and Citrus Red 2 would migrate through the gel at roughly the same speed, because they are the same size. Blue 1 and Fast green FCF would also move together.

Dog food manufactures might put food coloring in their food, because if a dog thinks something looks weird, they wont eat it. The will only eat it if it looks appetizing.

Two factors that contribute to how far dyes moved is the length of the molecule, and the time we ran the gel for.

Electricity, and positive attracting negative pulls the dyes through the gel.

The fact the smaller molecules move faster allows the molecules to separate. If they didn't you would get on band across all the used lanes, but since they do you can tell relative lengths

If given molecules with molecular weights of 600, 1000, 2000, and 5000 daltons I would expect the 600 to go the farthest, followed by 1000,then 2000, and finally the 5000 would barely have moved.

Recombination DNA

The first step to making recombination DNA is finding a plasmid and determining which antibiotic it is resistant to. You also have to identify the gene you want to add, for us we used the insulin gene, and sequencing it and the area around it. You then figure our which restriction enzyme can cut the plasmid in one location, and the DNA in two places, as close to the gene as you can. A restriction enzyme is an enzyme that looks for a specific dna sequence, and then makes a cut there. We used Xma I because it cut the plasmid in one location and the gene in two locations that were the closest to the insulin gene. If we had cut the plasmid in two locations we wold have removed part of the plasmid DNA, which we did not want to do. You then than ad lygase to re-join the plasmid. Finally you re-insert the plasmid into a bacteria, and add the antibiotic you determined the plasmid was resistant to, for us it was tetracyline. This kills all bacteria except the one you modified. Finally, you let the bacteria multiply.

This is important in every day life, because it is used to create many vaccines and medicines, by making bacteria that can produce these products. This technology can also be used to create GMO foods, such as fruits that spoil more slowly.

New year's goals

I will improve my test grades by studying more for my tests.

• I will start by setting aide time to study.
• I will make this time free of distractions.
• I will tailor my studying to my personal study style(visual, and kinesthetic)
• I will  study for at least an hour before every major test by the next bio unit test.
• I will make my studying more effective by tailoring to my own needs, and by finals, i will be an effective studyer

I will make finals in one robotics tournament.

• i will do this be working on the robot more
• i will include others in the team more 
• i will work more seriously during the meetings
• i will focus at the tournament 

Unit 5 reflection

In this unit we learned about  how DNA gets used to create proteins, and how DNA is copied. To copy DNA it is first unzipped by the enzyme helicase, then the enzyme DNA polymarase adds the missing halves, which creates two full sets of DNA. To create proteins, DNA is first transcribed into mRNA by the enzyme RNA polymarase. The RNA is then processed to remove unnecessary parts(introns), and leave only the extrons. The mRNA then leaves the nucleus, and goes to ribosome where it is read in 3 base sections called codon, and used to create a protein. The RNA can be mutated, and there are 3 types mutations. Substitutions are when one base is swapped for another, Insertions are when a base is added, and deletion is when a base is removed. Insertions, and deletion are called frame shift mutations, and have more effect on the protein than a substitution because they effect every codon after the mutation. Finally we learned about gene regulation, which controls which genes are expressed. This is done with an operon that can have a represser attach to at, and block the RNA polymerase from copying the DNA. I am pretty solid on everything in this unit. I am slightly less sure about gene regulation, but overall this unit was one on my better ones.

https://upload.wikimedia.org/wikipedia/commons/1/11/Gene_expression_control.png

http://study.com/cimages/multimages/16/point_mutation_types.png

In this unit learned some new skills. In the DNA extraction lab, we had to create our own procedure, and were not told if we had the right procedure, and this taught me to confident in myself. I still want to know more about how DNA expression, and regulation works. I find it interesting that despite the same DNA in all cells, they are very different, and I would like to know more about how and why that happens.

Protein synthesis lab

The body produces proteins in two steps, first DNA is transcribed into mRNA by an enzyme. The mRNA then leaves the nucleus, and goes to the ribosome. It attaches to the ribosome, and the ribosome creates a protein. The ribosome reads the mRNA in the base codons, and adds an amino acid depending on the codon. Once the ribosome reaches a stop codon it stop adding amino acids, and the protein is finished.




https://upload.wikimedia.org/wikipedia/commons/1/11/Gene_expression_control.png


The type of mutation with the least effect was substitution, because it only had an effect on one codon, this mutation has a good chance of having no effect if tit occurs in the last base pair of a codon. Insertion, and deletion had the most effect, because they effect all the codons after the point where the mutation occurs. These mutations have the most effect if the are toward the beginning of an mRNA sequence.



http://study.com/cimages/multimages/16/point_mutation_types.png


To make a mutation with maximum effect I substituted one of the base pairs in the first codon. I chose this because it would change the start codon, which would prevent the ribosome from making a protein at all. This mutation has to occur in the fist codon for it to have a major effect, otherwise it's effect will be very minor.



https://upload.wikimedia.org/wikipedia/en/6/67/Different_Types_of_Mutations.png


There is a genetic disorder called progeria, which has the symptom of an increased rate of aging. This usually results in the death of the person with the disorder due to a stoke or heart attack by the age of 20.

http://api.ning.com/files/rij9YXbYphAbAgYkKuHnEMcN0EoYnljvCN3Na44C-XRZsDjNcAW7VPfQcTyhkR8Esf49kNsXhP6BMl0uPyvyWw__/1.HutchinsonGilfordProgeriaSyndrome.jpg?width=229&height=350

DNA Extraction Lab

In this lab we asked the question: "How can DNA be separated from cheek cells order to study it?" We claim that DNA can be extracted from a cheek cell through a simple procedure. First, you scrap the sides of you mouth your teeth, then you swish you mouth with Gatorade, and spit it out. This lets you spit some of your cheek cells out with the liquid, and since Gatorade is a polar liquid it begins to break down the cheek cell. You then add salt to allow the DNA to clump up. You than add soap and enzymes from pineapple juice to further break down the cell, and at the end you ad alcohol to make the DNA float to the surface

As seen in the picture above the DNA floated to the surface at the end of this procedure, which proves that DNA can be extracted from a cell using this procedure.

On possible error is that some people poured the alcohol in too fast, and it mixed with the Gatorade mixture instead of forming a separate layer. This would negate the effect of the alcohol and prevent you from seeing DNA. To fix this the procedure should say to from two layers.

Another error is that is you don't put enough alcohol, then you wont see DNA. This could be fixed by adding more alcohol. 

This lab was done to show how DNA can be extracted from cells, and then be retrieved. This related to DNA, and how it is in every cell in your body. This lab can be applied to the field of genetics. You can use a similar procedure to extract DNA and test it for certain mutations.