pGLO lab Conclusion

Shiv Paul Gupta

pGLO Observations , Data Recording & Analysis

1.

Obtain your team plates.  Observe your set of  “+pGLO” plates under room light and with UV light.  Record numbers of colonies and color of colonies. Fill in the table below. Plate Number of Colonies Color of colonies under room light Color of colonies under   UV light - pGLO LB - pGLO LB/amp Unclear amount, bacteria grew in different shapes and sizes, and mixed together. No clear colonies A light brown/beige, color of diluted agar A light brown/beige, color of diluted agar + pGLO LB/amp 8 colonies with the same shape, of a sun A light brown/beige, color of diluted agar A light brown/beige, color of diluted agar + pGLO LB/amp/ara 7 colonies that lit up with ray like branches. Similar to a sun figure. A light brown/beige, color of diluted agar Fluorescent green

2.	What two new traits do your transformed bacteria have?
	The first is the distinct growth pattern or formation of the bacteria. They grow in a starfish/sun pattern where there is one main body of bacteria, and they more bacteria branches out from the main body. The second new trait is the fact that the main body of bacteria glows a fluorescent green due to the GFP fluorescent protein and the arabinose trigger.
3.	Estimate how many bacteria were in the 100 uL of bacteria that you spread on each plate. Explain your logic. Considering the miniscule size of bacteria, I can hypothesize that there are about 100,000,000,000 bacteria in 100 ul. I got this number because the average bacterium is about 0.2um, and doing the rough conversion of ul to um3I got my answer.
4.	What is the role of arabinose in the plates? Arabinose is used as a trigger for the plasmid. As we learned in the vodcast, the plasmids have two proteins. One is GFP, extracted from jellyfish. This protein is used to make the bacteria that picked up the plasmid glow. However, the GFP cannot do this on its own. It needs a trigger, which is where the arabinose sugar comes into play. It is necessary for the plasmid to actually glow.
5.	List and briefly explain three current uses for GFP (green fluorescent protein) in research or applied science. One application for GFP is its use for marking gene expression. Due to the lack of need for exogenous substrates and cofactors for this fluorescence, GFP expression can be used to monitor gene expression and protein localization in living organisms. Another application for GFP is like this lab, where it makes it easier to locate and differentiate bacteria from one another. A third use for GFP is to light up parts of the body for medical or scientific study. They can be used to highlight synapses in the body. This can make it easier to see and study.
6.	Give an example of another application of genetic engineering. One major use of genetic engineering, or genetic modification, is its ability to create and mass produce drugs and medicines such as insulin and growth hormones. The genetic engineering has lead to growth hormones that have treated dwarfism and other minor genetic defects.

Candy Gel Electrophoresis Lab Conclusion

When we analyzed the results of the gel, there were two dyes that had a different result than that of the reference dyes. The first dye that differentiated from the reference dyes came from Eng's candies, which were purple skittles. The original dye that came off the skittles was purple. However, at the end of the actual electrophoresis lab, the dye had separated into the same purple dye, but under the purple was a dark red dye. This was the exact color dye of the red reference dye. The red experimental dye that came out of the purple dye also traveled the same amount as the red reference dye. The photo below faintly shows the color separation in the purple experimental dye. The purple is five dyes from the left.

The second case of a differentiating dye came from my candies, the green Mike & Ike's. The green dye separated into a yellow dye that was identical to the yellow reference dye, and a blue dye that was identical to the blue reference dye. There was a significant difference between the distance traveled of the two dyes, but there were the same distances as the identical reference dyes. The photo above also shows this separation. It is the farthest right in the photo. 

The Fast Green FCF dye would travel similarly to the Blue 1 reference dye because their structure is virtually the same. The only difference is the molecules that make up the dyes. Citrus red 2 would travel a similar distance to both Yellow 6 and Red 40. This is not only because of the structure, but also because of the molecules that make the dyes up. Citrus red 2 is made up of OH, N, and O, while the two reference dyes are made up of HO, N, SO3, and O3S. There are only small differences in the makeup. Neither Carminic acid nor Betanin are similar to the reference dyes. 

Dog food manufacturers put artificial food dyes in the dog food to attract the owners of the dogs. Dogs that eat the food do not care about the color of the food they eat, considering the fact that they are color blind. However, their owners are conscious about what their beloved pets eat. It is just like buying food for their child, which is why they go for the most attractive foods and want what looks best for their pets.

Instead of keeping a record of our weekly intake, we decided to monitor the artificial dyes in popular foods. For example, fruit flavored, whole grain cereals considered healthy often had a mix of Blue 1 and Red 40. Specifically, fruit flavored Cheerios contains Red 40, Blue 1, and Yellow 6. It was also recently discovered that 19 of the most popular yogurt brands made their yogurt with Red 40. 

The two factors that most affect the distance traveled by the dyes are the charge and the length/size of the dye. The charge of the dye affects the distance because gel electrophoresis works on the DNA's electricity. The DNA is attracted to positive charges, which means that charge of the DNA/dye tested matters. The size/length of the dye matters because the smaller the strand, the faster/farther it travels. 

Electricity is the force that helps more the dyes through the gel. The dye is attracted to the positive charge at the other end of the gel .

The electricity component also causes the molecules to separate by size. This is because the dyes move across the gel plane with the positive charge. However, the dyes move a certain distance in a certain amount of time. The size affects the distance traveled because the larger the dye, the longer it takes to get from one place to another. It is like a small Porsche racing against a large 18-wheeler. The smaller vehicle will win because it is smaller and there is less mass to move from the start to the finish. 

Size corresponds to weight. Mostly, the larger something is, the heavier it is. Going by the logic stated in the previous paragraph, that the smaller/lighter the object the farther it moves, the fewer daltons a DNA strand weighs, the farther it will travel. The DNA that weighs 600 daltons will travel the farthest, while the 5000 dalton DNA will travel the shortest distance. Everything else falls in between.  

2017 New Year's SMART Goals

2016 was one of my best years yet. Not just because of grade, but also because of obstacles I have overcome from family, to friends, to school. An example of this was when my sister Shazia went to college. The day she left, everyone was excited for her but sad at the same time. However, I was just excited. I did not think I would miss her very much. As time went on with an empty, lonely bedroom in the house, I began to feel it. I would turn the corner of the staircase to her room and look inside, intending to annoy you, but then I would realize nobody was there. It was only after she left that I realized how much I interacted with her and missed her. When she came home for winter break, I could not spend enough time with her. She is an amazing sister.

Another example of family occurred over December break. Maya and I don't usually get along very well, and we would constantly get into fights over the littlest things. However, as if a whole other person took control of her body, Maya was one of the best older sisters anyone could ask for. She was there for me when I was sick, when I injured my arm, and just to hang out. It was crazy how all of a sudden she had changed. I had tried to mend our relationship many times before, but I was not great at being "the bigger man".

These changes and accomplishments last year lead to a new found determination to improve myself. One thing I noticed in this class especially is my lack of determination and punctuality in the class. I did not take this class seriously at first because I did not like the teaching method of lectures at home. When there was a blogger post or a vodcast due, I would usually do them at home, but then sometimes just blow it of because I was tired. Considering the fact the I play a sport now, I have a free fourth period. However, my 2017 new year goal is to do the homework at home, and stop procrastinating. This is more of a mental goal. One thing I am going to do is put my phone away during the vodcast. That is a major distraction for me. Another thing I am going to do is set a specific time for myself daily, or every other day, on when to do biology. Although I should have done these things in the first place, I did not, but I will do them now.

My second new years goal will be to get to school on time everyday for the rest of high school. I would usually show up to school about 2-5 minutes late about 2-3 times a week, which is surprising, considering the fact that I was punctual in middle school. The main reasons for my tardiness are lack of sleep and an increase in traffic to get to SHS. My plan this year is to finish my homework by 9-9:30pm latest (putting away all distractions such as my phone will help), and to be more productive while getting ready in the mornings. This will not only give me more energy, but then I will be able to improve in school as a whole.