Gel Electrophoresis

So what exactly is this gel electrophoresis thing anyway? Well being that DNA molecules are so small that you can't see them in most microscopes, in order for people to see what DNa actually looks like, they came up with this amazing thing called gel electrophoresis. Most scientiest use this for sorting DNA strands by length. The gel is a sponge like thing made of Jell-O that has many small holes in it. The electrphoresis is how the DNA is pushed through the gel filter. The electric current makes the DNA move. Small strands tend to move faster, and the longer they go the further they get from the starting point. Longer ones obviously tend to move slower. Strands that are the same size sometimes stick together. To make the DNA visible to the human eyes, they must be stained. Even with them being stained, we still can't see single strands of DNA, only larger groups.

DNA Sequencing!!! :)

DNA Sequencing Data!!!

While going through the DNA activity, an importance was to see which patient's sequence matched that of a normal one. In doing so, it was found that Abby and Bob had the most similar sequence to the normal. This would expect to have a normal state, which means most likely no diseases. Carol on the other hand had a very low percentage compared to the normal sequence. With this being said, she would most likely be more in the disease state. After review the sequences together, it was found that in Carol's DNA a letter was skipped somewhere along the way making everything else in her DNA shift. Just the slight factor of a letter being gone made a huge difference in this case for Carol.

Transformation Lab

Steps to the transformation lab:

1.) The first step was to get organized, and label all of our material. We labeled a green tube with a (+) for +pGLO and on orange tube with a (-) for -pGLO. The tubes were then placed into a foam rack, until needed. 

2.) Having both the green and the orange tube open, we used a sterile transfer pipet to transfer 250 ul of (CaCl2) also known as the transformation solution. After both tubes being filled with the transformation solution, the tubes were placed on ice to prepare for the next step.

3.) We then used a sterile loop to pick up a colony of bacteria from a starter plate. We placed an unknown amount of colonies in both the green (+) and the orange (-) tubes. 

4.) Soon after, we found ourselves waiting for the plasmid DNA solution. With a new sterile loop, it was a difficult task to get the plasma to fill the loop. Once it was finally filled, we inserted the plasma into the green tube (+) but not the orange tube (-). 

5.) The tubes were then placed in ice for 10 minutes. While we waited for the tubes on ice, we labeled our 4 agar plates with: LB/amp (+), LB/amp (-), LB/amp/ara, and LB.

6.) The tubes were then placed into the foam rack and placed into heat shock of 42 degrees for 50 seconds. After the 50 seconds were up, the tubes were placed back into the ice container for 2 minutes. 

7.) Once the 2 minutes were up, we then placed 100 ul of LB nutrient broth in both tubes. The tubes were then incubated for 10 minutes at room temp. 

8.) After the 10 minutes were up, we then used a sterile pipet for each plate, and added 100 ul of transformation and control suspensions into the appropriate plates. We then spread the suspensions evenly over the surface.

9.) The plates were then stacked together, and the stack was placed in the incubator at 37 degrees until the next day.

Expectations:

We expected to find the most bacteria on the LB, because it wasn't tampered with as much. We decided that the LB/amp and the LB/amp/ara would have genetically transformed bacterial cells, due to the fact that they have antibiotics. The +pGLO amp and the -pGLO amp we decided should be compared to determine if there was any genetic transformation that occurred. 

Results:

We drew what we saw on each plate, and placed our information into a data table. The -pGLO LB/amp came up with no colonies. The +pGLO LB/amp we found 2 colonies. The +pGLO LB/amp/ara had 6 colonies. The -pGLO LB had 8 colonies.

Gattaca Movie Review!

Here is a link to a glogster talking a little bit about what I thought of Gattaca.

Gattaca!!!

In the movie Gattaca, its a space agency that is supposed to be in the near future. At this agency, they are planning a trip to Saturn's moon Titan. In order to take the journey, you must have the right genetic makeup. Vincent Freeman wants badly to make the trip, but in Gattaca he is considered an "in-valid", which means he has defective genes. Vincent "switches" places with Jerome Morrow, right down to the nucleic acid. Vincent takes hair, blood, and urine samples from the real Jerome, to pass all of the required tests at Gattaca. Due to the fact that the real Jerome Morrow was crippled in some kind of accident, Jerome needs Vincent just as much as Vincent needs Jerome.

The scientific premise, like in most science-fiction movies, combines a mix of truth and fiction. In the movie, the alleles from parents are so chosen that the combination produces the optimal arrangement in terms of the child's genotype. But we know enough right now to realise that even in situations where there is a great degree of genetic predisposition, it is quite probable that that predisposition (positive or negative) is never realised. For more complex behavioural traits such as intelligence, aptitude test results would be a far better indicator than genetic makeup. In other words, any correlation people may find between a complex behavioural trait such as intelligence and genetics is for all practical purposes controlled by the environment, given the "edge of chaos" nature of such traits.




"What is disturbing about our genetic engineering capabilities today is no more disturbing than our medical engineering capabilities (and there are plenty of disturbing ramifications). The genetic component simply provides one additional way to discriminate in the real world, just as it is routinely done with age, sex, years of experience, education, and physical ability: Consider the physical and mental requirements for being an astronaut today, or even for admission to college. Information, of any sort, is a valuable commodity in this day and age, and the kind encoded in DNA is no exception. Humans naturally use information to discriminate. I would argue that for some of scenarios posited in Gattaca, the genetic information is far less reliable than physiological and psychological histories. "

Information from:
http://www.ram.org/ramblings/movies/gattaca.html

Bartonella Henselae

While doing a virtual lab on bacterial DNA, I was able to be a lab technician in a molecular biology lab and was responsible for providing lab results for use in diagnosing patients. On the certain DNa that I found, it came up with the bacteria Bartonella Henselae. Here is what was found:
Bartonella henselae Various species of Bartonella that are pathogenic to humans are transmitted via a vector, or directly from an animal reservoir. For example, B. bacilliformis via sandflies causes Oroya fever; B. quintana via body lice causes trench fever; and B. henselae via cats causes cat scratch disease (CSD). CSD typically manifests as swellings of the lymph glands, possibly with skin lesions at the site of inoculation and possibly accompanied by fever, fatigue, and other symptoms. Immunocompromised patients may be particularly susceptible and can develop a different disease, bacillary angiomatosis, as a result of infection by B. henselae or B. quintana.

Below are pictures of Bartonella Henselae:

How I was able to sequence the DNA:

In the first step, my objective was to:

• Prepare a sample from a patient and isolate whole bacterial DNA.
• Make many copies of the desired piece of DNA.
• Sequence the DNA.
• Analyze the sequence and identify the bacteria.