Tuesday, May 10, 2011

DNA Fingerprinting

DNA Fingerprinting is so extremely interesting to me. Each and every person has their own set of fingerprints, and none are ever the same. Even identical twins have different fingerprints even if they are semi-similar. Both are hands and fingers have different shapes and lines of the indentions of each person which is what makes us all so different from one another. With the advancements of today's technology scientists are able to identify almost anyone with just the slightest part of a fingerprint or handprint. Fingerprinting has come a long way and is used to identify fugitives everyday. Fingerprints are now the greatest lead to identifying and prosecuting someone who has committed a crime. It is even recommend that once your child has reached an age where they can walk on their own it is important to get a home kit to take your child's fingerprints and other information just in case something bad happens to them. These little things can make and break a case in whether the police can identify your child or not. In crime shows and in real life dusting for fingerprints is one of the first things done to a crime scene. This is the initial lead to all cases.
The identification of fingerprints is known as dactyloscopy. Dactyloscopy is a process of comparing two instances of skin impressions. Human fingers, the palms of your hand, even your toes can determine if the impression came from the same person. Friction ridge is the form in which prints are left, and not even the same person's fingerprints, handprints, or even footprints are exactly the same every time. The smallest of details change with each touch and movement. Just as i am typing right now. Each key I have used has my finger print on it, but each print is different in some minute way. Fingerprint identification is also referred to as individualization. A computer system had been built and edited and revised constantly in order to accurately be able to identify specific fingerprints of the persons in the data base. These computers are operated under a threshold scoring, which is the process of determining whether or not two impressions are from the same person or not. Intentional recording of fingerprinting is typically taken using black ink on a white paper, to show the contrast. The finger is pressed down then rolled to be sure that the entire print is taken. A live scan is done on fingerprints that are invisible to the naked eye, but can be seen with chemical methods and powders. These types of prints are called "latent prints."
So overall fingerprints vary in many ways whether its from size, shape, or the clearness of the impression. So always remember if you plan to commit a crime you can and will be identified with just the slightest of impression left behind. ;)
Link

Electrophoresis!!!


Hey look its Sidney and I using the rather large pipets used in creating this gel electrophoresis lab! Freeze! Nobody move!!






This little contraption to your right is the gel rig where the gel slab sits during the experiment.








Here is a picture of a gel slab in the making!!!










In this Gel Electrophoresis lab I learned many new lab techniques throughout this rather intersting process. The overall point of this lab was to insert different colored dyes into the gel slab and then rn electrical currents through them to see how and what type of change the dyes would go through. The first step in this lab was to make one of the get slabs that had to set and rest for around twenty minutes. To create the gel slab you needed to make a mold using the intersecter things to create the rectangular shapes where the dyes would be placed. Then you filled the tray up with melted gel liquid being sure it is as even as possible all the way across. After the gel has set we placed it in a holder for lack of a better word. From there Mr. Ludwig taught us how to use the pipets and the dyes that we would be injecting into the gel slab. With each new dye the pipet nozzle had to be changed to avoid contamination. Our groups then began this process of adding dyes to each tiny individual slot. This part was a little challenging because of how small the squares were. After the slots had all been filled we added the electrical current to the experiment and let it sit. By the time we came back to school the next day all the negative dyes had moved in the direction of the positive dyes, whereas the single positive dye moved towards the negative side. Each of the different dyes seemed to travel different distances across the gel slab, depending on how the dye was changed. Overall this process was very fun and intriguing and contributed to my understanding of Gel Electrophoresis and how it is used in DNA sequencing.

Bacterial Transformation

Bacterial Transformation and its Process
Step #1: You need to label one of the closed test tubes +pGLO and another -pGLO, then place it in the foam tube holder.
Step #2: Open the tubes and use a pipet to transfer 250ul of the CaCl2. Place this tube on ice.
Step #3: Use the sterile loop to pick up a single form of bacteria from the starter plate. Get the +pGLO tube ready, then immerse the loop into the transformation solution at the bottom of the tube. Spin the loop until the entire colony is dispersed. Place the tube back in the ice bucket. Repeat with -pGLO with a new sterile loop.
Step #4: Examine the plasmid DNA with the UV light. Immerse the new plasmid into the DNA stocktube. Withdraw another loop of bacteria. In this step there should be a film of plasmid across the ring. Mix the loopful into the cell suspension of the +pGLO tube. Once again put it back on the ice. DO NOT add plasmid DNA to the -pGLO tube.
Step #5: Incubate the tubes that are on ice for at the most 10 minutes. The bottoms of the tubes should be in contact with the ice.
Step #6: Next label four agar plates as follows: LB/amp plate label +pGLO; LB/amp/ara plate label -pGLO; LB/amp label -pGLO; and the LB plate -pGLO.
Step #7: After that step transfer both tubes into the water bath that's set at 42 degrees Celsius for exactly 50 seconds.
Step #8: Place the tubes in ice for two minutes then take them off the ice and open the tube. Use a new pipet to add 250 ul of LB nutrient broth to the tube and reclose it. Repeat with a new pipet to the other tube. Incubate the tubes at room temp for 10 minutes.
Step #9: Mix the tubes softly. Use a new sterile pipet for each tube, pipet 100ul of the transformation control suspensions to appropriate plates.
Step #10: Spread the suspensions evenly with a sterile loop.
Step #11: Stack the plates and tape them together, upside down. Place it in the Biology incubator overnight which is set around 37 degrees Celsius.

Observations:
+pGLO; LB/amp: has the gene but it isn't "turned on". It has a medium to heavy population which is white and yellow. Our estimated count was 130. Transformation plates
+pGLO; LB/amp/ara: has the protein that is turned on, which makes it glow. It has a medium to heavy population that's yellow then a light white. Our estimated count is 149. Transformation plates
-pGLO; LB/amp: has no population and no bacteria!! Control Plates
-pGLO; LB: has a HEAVY bacterial lawn!!! Control Plates

Overall Analysis:
Plasmid seemingly does not make a difference to the health of the illness the bacteria may cause.
The amp kills the bacteria unless a gene from the plasmid is LB/amp +/-.

Friday, May 6, 2011

GATTACA!

GATTACA Movie!!! =)
I wasn't here for most of this movie, therefore I kinda had to wing it, while using my other resources. But I tried! =)

Questions Regarding GATTACA!
1. The following terms were used in the movie. How do they relate to the words we use: degenerate and invalid?
         De-gene-erate: What a god child is called
         In-valid: The type of person that wasn’t made
         Borrowed Ladder: A person that you disguise yourself as

2. Why do you think Vincent left his family, tearing his picture out of the family photo, after winning the swimming race against his brother?
I think he finally left the family so he could start his life over in a sense and prove his point to his brother that he could do whatever he set his mind too even though he was different. However after he finished that he seemed to have the strength to move on with his life, and finish things he thrived to do.

3. Describe the relationship between Vincent and Anton.
Vincent and Anton, were brothers in this movie. They were extremely competitive with one another. Both of the two were always trying to be better than the other at multiple things. Anton just happened to be the detective in Vincent's case which added to this sibling rivalry.

4. When Jerome Morrow said to Vincent/Jerome, “They’re not looking for you. When they look at you, they only see me,” what did he mean? Can you find any parallels to this type of situation in real life?
 What he meant by this situation was that technically speaking Vincent no longer existed now that Jerome had become what Vincent once was. Therefor no one realized that Jerome was around because the connection between the two was never made. Also this conversation could have meant that being accused of things would no longer happen to Vincent because his record show him as another person. I would say that the parallel in this situation is when Jerome hears something about his old self. He would more than likely become nervous because he is still the same person overall.

5. Choose your favorite character from the film. Explain why you choose that person. Would you want to be that person? Why? Why not?
My favorite character in GATTACA would have to have been Vincent. In my opinion he was the most prominant character with an idea and a mission, which added to the excitement of his character. Also he was very determined to accomplish certain activities and that drive was a great characteristic to me. However I did feel bad for him since he had to change his identity. 

6. At the end of the film, you are told that the Doctor knew about Vincent all along. Why did the Doctor go along with the fraud? What would you have done if you were the Doctor?
            I would have to say that the doctor went along with the deception the whole time because he was on Vincent's side and he wanted him to accomplish what he set out to do. Personally I would probably have went along with it as well because I wouldn't want to rat someone out that had worked so hard to be an achiever.

7. The technology to do what was done in the movie is definitely possible within the next fifty years. Do you think that Vincent’s world could eventually happen in America? Why?
 I don't think this would necessarily happen in America because we have rights within the government. I'm sure throughout the years this process could happen, especially with how fast our technology changes, so I wouldn't doubt it occurring, but I don't believe it will be that popular in America.

8. What do you think is wrong with the society portrayed in "GATTACA"? What is right?
 I don't believe anything was necessarily right in this movie. The whole point of the parents choosing how the baby would be was in my opinion wrong. You should love the baby you conceive without changing it. The discrimination that was presented against the "invalid" ones, was ridiculous to me. After all nothing is perfect. To me it seemed like if you weren't perfect than you weren't good enough and I really disliked that situation.

9. What were the screenwriters trying to tell us through the episode of the 12-fingered pianist? Is anything wrong with engineering children to have 12 fingers if, as a result, they will be able to make extraordinarily beautiful music?
 I believe the screenwriters were trying to say that people would start trying to engineer children to be good at certain things and succeed in them, which is obviously wrong because its what the  parents want not the children.

10. You and your spouse are having a child and are at the Genetic Clinic pictured in the movie. What characteristics would you want for your child and what would you ask to be excluded? Why would you make those choices?
Normally I woul say that there is no way I would go to a clinic sugh as this because of the ways they discriminate against babies that are yet to even be born. But if I had to choose I would want them to be very athletic, tall and smart. However I would take out my families "mole" gene from the sequence.

11. Picture yourself as either Vincent, Jerome, or Anton. Would you have acted the same or done things differently if you were in the same world as them?
 I probably would've acted the same as they all had. Each of them handled each situation in the best way that they could and I don't think anyone should doubt them about their actions. However I would not have committed suicide like Jerome did.


12. How does the society in GATTACA resemble the type of society America was during the height of the eugenics movement?
People were extremely anxious and excited about being able to create their own baby and help make them "better" in their eyes which is just like in GATTACA. However in GATTACA the clinic babies were the only ones who were accepted for the eugenics experiment.

Overall I enjoyed the parts and pieces I seen to the GATTACA movie. It was a very different movie, but it taught me a lot about eugenics. However I hope America never turns into a country where we create our children instead of just conceiving them naturally.

Monday, April 25, 2011

PCR!!! Yay!

Online Bacterial Lab

Doing this online Bacterial ID Lab was very helpful, it provided a procedure of identifying bacterial samples that were given.
The Polymerase Chain Reaction Machine, better known as The PCR Machine is an expensive machine that is worth thousands of dollars. The PCR Machine provides information about DNA sequencing and is very important to this process. The work that a PCR machine conducts is altering the temperature of DNA. This type of machine takes a strand of DNA, and adds heat to the situation to in a sense "Melt" the DNA. It separates the DNA into two single strands from its double helix view. From there the machine cools the separated strands and each strand is primed. After the DNA strands have been stabilized to room temperature the primer copies a segment of DNA that is specified before hand.
 So overall the PCR Machine is a really important and helpful attribute to DNA sequencing, it helps in variations of defect detecting and even DNA TESTING!!

Now for The LAB!
-In this ID Lab you had to prepare DNA samples that were going to be used in the experiment. This step involved placing the bacterial colony in a microcentrifuge tube and inserting the digestive buffer. After that you have to heat the inactive digestive enzymes, which will denature and inactivate the enzymes. From there you add a counterbalance and add cell debris for removal of the sample. After these steps are completed the DNA becomes supernatant. Transfer the supernatant to the PCR tube and your prepping is done.

-The next step is to use PCR to make copies of these DNA sequences. In order for this to  happen you have to add a PCR Master Mix solution for all of the tubes. To set up a positive control reaction, you must add controlled DNA to one of the tubes. However, for negative reactions you must combine de-ionized water to one of the tubes as well. After that you must insert both the tubes into the PCR Machine. The short strands that are extracted are the DNA strands that were to become of the solution from the PCR thermal cycling process.

-Continuing on from there you must purify the PCR solution. In order to do this step you have to set up the microconcentrator column and add a buffer solution to the column, then add the PCR solution to the column. The positive and negative controls will be placed in ice until they will be needed again. Then add counterbalance to the tube and put it in the centrifuge. After time has passed the DNA will have become stuck into the column and you must loosen it and put it into another tube separate from the others. Place the column into the new tube and add a buffer. From there you must add a centrifuge to be able to collect the DNA.

-The next step in this process is the PCR sequencing. First you must dilute the PCR product to increase its overall volume. Add distilled water to the new PCR Product in order to purify it. The blue and green strip tubes contain PCR reactant mixtures. Now you need to add the purified PCR solution to the mixtures. They are now ready for the PCR Amplification Process. Place the tubes into the PCR Machine. This step is now finished as well.

-Next you must load the auto sequencer and insert the tubes into it. In my first testing in this ID Lab my DNA sequence read tacggtagcgtaatgcc. Overall that completes the DNA sequencing.

Thursday, March 31, 2011

DNA Sequencing

In this experiment we used the Sanger Method of sequencing DNA. Or in other words the older version of sequencing DNA, considering that since this time we have advanced in technology and sequencing DNA has become easier and faster to be done. In the Sanger Method the letters are written down in threes in the order that they occur in the sequence. The groups of threes are placed and separated, then the selection of the protein begins that corresponds with the groups pattern. Abby was the first patient and she had only one letter difference, thus meaning she had a point mutation or a single base change. Bob was the next patient, he also had one difference in his DNA sequence. However with the type of protein sequence he acquired his sequence stopped short which lead to him having a truncation mutation. The last patient was Carol she had the most DNA sequencing differences from the normal sequence. Carol  had a frameshift mutation. A frameshift mutation is where there is a gain or loss of a letter that throws the rest of the sequence off from its normal pattern. So although we all have DNA and it has its own sequencing pattern there are plenty of mutations that can occur and cause the sequence to mess up or end overall.







  That is how you read a sequence and the three things that can go wron